A Phase 1 Study of M032 (NSC 733972), a Genetically Engineered HSV-1 Expressing IL-12, in Patients With Recurrent/Progressive Glioblastoma Multiforme, Anaplastic Astrocytoma, or Gliosarcoma
Phase: PHASE1
Conditions: Recurrent Glioblastoma Multiforme, Progressive Glioblastoma Multiforme, Anaplastic Astrocytoma or Gliosarcoma
Sponsor: University of Alabama at Birmingham

 
 Phase 1 Study of M032 (NSC 733972), a Genetically Engineered 
HSV-1 Expressing IL-12, in Patients with Recurrent/Progressive 
Glioblastoma Multiforme, Anaplastic Astrocytoma, or 
Gliosarcoma 
 
Study Protocol & Statistical Analysis Plan 
 
NCT02062827 
 
 
Date of Last Protocol Approval 
10 August 2023 
 
 
James Markert, MD  
University of Alabama at Birmingham 
Birmingham, AL 35294 
 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 1 of 66 
Version 2.9 dated 20 Sep 2021 
 
 
 
TITLE:  A Phase 1 Study of M032 (NSC 733972), a Genetically Engineered HSV-1 
Expressing IL-12, in Patients with Recurrent/Progressive 
Glioblastoma Multiforme, Anaplastic Astrocytoma, or 
Gliosarcoma 
 
Version: Amendment 12 Version 2.9 Date: Sep 20, 2021 
Version: Amendment 11 Version 2.8 Date: Jan 20, 2021 
Version: Amendment 10 Version 2.7 Date: Aug 25, 2020 
Version: Amendment 9 Version 2.6 Date: Apr 6, 2020 
 Version: Amendment 8 Version 2.5 Date: Dec 10, 2019 
 Version: Amendment 7 Version 2.4 Date: Apr 5, 2019  
 Version: Amendment 6 Version 2.3 Date: Feb 22, 2019 
 Version: Amendment 5 Version 2.2 Date: Jan 15, 2019 
 Version: Amendment 4 Version 2.1 Date: May 30, 2018  
 Version: Amendment 3 Version 2   Date: April 28, 2017 
 Version: Amendment 2 version 1.8 Date: August 26, 2015 
 Version: Amendment 2 version 1.7 Date: April 20, 2015 
 Version: Amendment 1 version 1.6 Date: July 22, 2014  
 Version: Revision 1.5 Date:  August 20, 2013 
 Version: 1.1 Date: August 20, 2013 
 Version: 1.0 Date: December 15, 2011 
 
SPONSOR: 
NIH/NCI 
 
IND#:   
14,946 
 
NIH Grant: 
1R01CA217179 
 
NCI Supplied Agent: 
M032 (NSC 733972, supplied by NeXT program  
 
Coordinating Center: 
 
Single-site trial at the University of Alabama at Birmingham 
 
Principal Investigator: 
James M. Markert, M.D.  
 
Professor and Chair, Department of Neurosurgery 
 
The University of Alabama at Birmingham 
 
FOT 1060 
 
1720 2nd Avenue South 
 
Birmingham, Alabama 35294-3410 
 
Phone: 205.934.7171 
 
Fax:  205.934.9820 
 
Co-Investigator:  
 
 
L. Burt Nabors, M.D. 
 
  
 
 
 
 
510 20th Street South, 1003 FOT  
 
 
 
 
 
  
 
 
 
 
Birmingham, AL 35294-3410 
 
 
 
  
 
 
 
 
Phone: (205) 934-1813 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 2 of 66 
Version 2.9 dated 20 Sep 2021 
E-mail address: bnabors@uab.edu 
 
 
Statisticians:   
 
 
 
Inmaculada Aban, Ph.D. 
 
 
 
Gary R. Cutter, Ph.D. 
 
 
 
 
 
 
 
 
Department of Biostatistics  
 
 
Department of Biostatistics 
 
 
 
 
 
 
 
 
1720 2nd  Ave South , RPHB 414 
 
1720 2nd Avenue South, RPHB 327 
 
 
 
 
 
 
 
 
Birmingham, AL 35294-0022 
 
 
Birmingham, AL 35294-0022 
 
 
 
 
 
 
 
 
Phone: (205) 934-2732  
 
 
 
Phone:  (205) 975-5048 
 
 
 
 
 
 
 
 
Email address:  caban@uab.edu   
 
E-mail address: cutterg@uab.edu 
 
Responsible Research  
 
Mary Jane Avant, RN  
Nurse Coordinator: 
 
 
510 20th Street South, 1003 FOT 
Birmingham, AL 35294-3410 
Phone (205) 934-8347 
Fax (205) 975-9928 
 
 
 
 
 
 
 
 
E-mail address:  mavant@uabmc.edu 
 
Responsible Data  
 
 
Hui-chien Kuo  
Manager: Department of Biostatistics 
1720 2nd Ave South 
 
 
 
 
 
 
 
 
Birmingham, AL 35294-0022 
 
 
 
 
 
 
 
 
Phone: (205)   975-9247 
 
 
 
 
 
 
 
 
Email address:  huichien@uab.edu 
 
 Research Pharmacy:  
 
UAB Research Pharmacy 
UAB Hospital Investigational Drug Service Pharmacy 
1802 6th Avenue South, NP 3470 
Birmingham, AL  35249 
Phone: 205-934-7191 
Fax: 205-975-6647 
 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 3 of 66 
Version 2.9 dated 20 Sep 2021 
Changes between Amendment 11 version 2.8 dated 20 Jan, 2021 and Amendment 12 version 2.9 dated 20 Sep 2021 
 
Section/location 
Change  
Rationale 
8 
 
Adding an unscheduled 
visit 
 
Administrative change 
Study synopsis 
 
In a previous amendment it 
was stated that 4-20 
participants are to be enrolled 
and treated in addition to the 
first 4 in the seed model 
(Section 4.1.2). The 
enrollment number is being 
increased to 24 to clarify that 
the previously stated 4-20 
participants are in addition to 
the first 4. 
Administrative change 
 
 
 
 
 
 
 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 4 of 66 
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STUDY SYNOPSIS 
  
 
Objective: 
To obtain safety information in small cohorts of individuals (three patients per 
group), with cohorts to receive escalating doses of M032.  Safety will be assessed 
at each dose level before proceeding to the next dose.  Biologic secondary 
objectives include characterization of the in situ activity of M032 after 
intratumoral inoculation and of the local and systemic immune responses to M032.  
As a clinical secondary objective, patients will be followed serially by MRI for 
potential clinical response to M032. The clinical strategy takes advantage of the 
virus’ ability to infect and lyse tumor cells and the potential for enhancement of 
this effect by the induction of an anti-tumor immune response by IL-12 as well as 
an anti-angiogenic response by this cytokine. 
Treatment Indication: 
Progressive growth of glioblastoma multiforme, anaplastic astrocytoma or 
gliosarcoma after radiation therapy. 
Clinical Phase: 
Phase 1 (open-label) 
Design: 
 
 
 
 
Single dose of M032 infused through catheters into region(s) of tumor defined by 
MRI; Up to 3 patients/dosage level; dosage escalation proceeds only after a 
minimum of 24 days of observation, if incidence of Grade III/IV toxicities is 
acceptable.  Dose increases or reductions will be determined using a modified 
Continual Reassessment Method (CRM); extent of dose changes for subsequent 
subjects will be increased up to, but not exceeding the next higher dose level (1 
log). Dose modifications will utilize a modified CRM for each successive subject 
until an MTD or the maximal planned dose is reached. 
Study Duration Per Patient: 
12 months 
Subject Population: 
8 to 24 patients with recurrent/progressive glioblastoma multiforme, anaplastic 
astrocytoma or gliosarcoma depending on toxicities.  (Numbers may vary 
slightly pending acceptance of previously treated patients to update the CRM) 
Study Medication and 
Dosage: 
Dose escalations of up to, but not exceeding the next highest dose level (1 log). 
Dose escalations from 1 x 105 to 1 x 109 plaque-forming units of M032, a 
genetically engineered herpes simplex virus type 1 expressing human IL-12 
(NSC 733972).  
Safety Evaluations: 
Follow-up evaluations using routine laboratory analyses and clinical 
measurements of neurological function and evidence of M032-related toxicity. 
Studies to evaluate the possibility of M032 shedding will also be conducted.  
Patients will be observed closely during the planned post-treatment 
hospitalization period, followed by outpatient evaluations done at 10 days, then 
months 1, 2, 3, 4, 5, 6, 9 and 12, subject to disease progression. 
Study Endpoints 
Primary:  CRM-estimated highest safe dose or maximal planned dose 
if no dose-limiting toxicity observed. 
Secondary: Time to progression, survival, biologic assessments. 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 5 of 66 
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TABLE OF CONTENTS 
 
Section 
 
 
Page Number 
 
SYNOPSIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .   3 
 
 
 
1. 
OBJECTIVES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
6 
 
1.1 
Lay Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  6 
 
1.2 
Primary Objective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
6 
 
1.3 
Secondary Objectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  6 
 
1.4 
Tertiary Objectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  6 
 
 
 
 
 
 
2. 
BACKGROUND . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
6 
 
2.1 
Safety and efficacy of non-virulent, oncolytic Δγ134.5 HSV . . . . . . . . . . . . . . . . .  
6 
 
2.2 
Malignant Glioma . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  10 
 
2.3 
Rationale. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 
 
 
 
 
 
 
3. 
PATIENT SELECTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  11 
 
3.1 
Eligibility Criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
11 
 
3.2 
Exclusion Criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  12 
 
3.2 
Inclusion of Women and Minorities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  13 
 
 
 
 
 
 
4. 
TREATMENT PLAN . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .   13 
 
4.1 
Study Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  . 
13 
 
4.2 
Definition of Dose-Limiting Toxicity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
18 
 
4.3 
Supportive Care Guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  20 
 
4.4 
Duration of Therapy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  20 
 
4.5 
Halting Rules for Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  20 
 
 
 
 
 
 
5. 
EXPECTED ADVERSE EVENTS/DOSE MODIFICATIONS . . . . . . . . . . . . . . . . . . .  
21 
 
5.1 
Expected Adverse Events Associated with M032 . . . . . . . . . . . . . . . . . . . . . . . . . .  
21 
 
5.2 
Dosing Delays/Dose Modifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
23 
 
5.3 
Adverse Assessment and management Assessment by PI/Clinician . . . . . . . . . . .  23 
 
 
 
 
 
 
6. 
AGENT FORMULATION AND PROCUREMENT . . . . . . . . . . . . . . . . . . . . . . . . . . . .  23 
 
6.1 
M032 Formulation and Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 
23 
 
6.2 
Availability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  . . . .  23 
 
6.3 
Excipient for Dilution of M032 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  24 
 
6.4 
M032 Dose Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  24 
 
6.5 
Precautions in Handling M032 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  24 
 
6.6 
Precautions in Disposal of M032 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  24 
 
 
 
 
 
 
7. 
CORRELATIVE/SPECIAL STUDIES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  24 
 
 
 
 
 
 
8. 
STUDY CALENDAR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
26 
 
8.1 
Study Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  27 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
9. 
MEASUREMENT OF EFFECT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
33 

NCI  Protocol #:  N/A 
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M032 Amendment 12 
University of Alabama at Birmingham 
Page 6 of 66 
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9.1 
Definitions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  33 
 
9.2 
Guidelines for Evaluation of Measurable Disease . . . . . . . . . . . . . . . . . . . . . . . . . .  33 
 
9.3 
Response Criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  34 
 
9.4 
Confirmatory Measurement/Duration of Response . . . . . . . . . . . . . . . . . . . . . . . .  35 
 
9.5 
Progression-Free Survival . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  35 
 
 
 
 
 
 
10. REGULATORY AND REPORTING REQUIREMENTS . . . . . . . . . . . . . . . . . . . . . . .   35 
 
10.1 Expedited Adverse Event Reporting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  35 
 
10.2 Data Reporting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
37 
 
10.3 CTEP Multicenter Guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  37 
 
10.4 Cooperative Research and Development Agreement (CRADA)/Clinical  
Trials Agreement (CTA) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
37 
 
 
 
 
 
 
11. STATISTICAL CONSIDERATIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  37 
 
11.1 Study Design/Endpoints . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  38 
 
11.2 Sample Size/Accrual Rate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
38 
 
11.3 Stratification Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
38 
 
11.4 Analysis of Secondary Endpoints . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
38 
 
 
 
 
 
 
12. REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  39 
 
 
 
 
 
 
APPENDIX A:  Expected Adverse Events Associated With M032 . . . . . . . . . . . . . . . . . . . . . . .  42 
 
 
 
 
 
 
APPENDIX B:  Performance Status Criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
43 
 
 
 
 
 
 
APPENDIX C:  Instructions for M032 dose preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
44 
 
 
 
 
 
 
APPENDIX D:  M032 dose preparation guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  46 
 
 
 
 
 
 
APPENDIX E:  M032 shipment information/handling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  50 
 
 
 
 
 
 
APPENDIX F:  Serious Adverse Event Reporting Form (MedWatch)  . . . . . . . . . . . . . . . . . . . .  51 
 
 
APPENDIX G:  Data and Safety Monitoring Board . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .  
64 
 
 
 
 
 
 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 7 of 66 
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1. OBJECTIVES 
1.1 Lay Abstract 
 
M032 is a second-generation oncolytic herpes simplex virus (oHSV) that is conditionally replication 
competent; that is, similar to G207, a first generation oHSV, it can replicate in tumor cells, but not in normal 
cells, thus killing the tumor cells directly through this process. Replication of M032 in the tumor itself not only 
kills the infected tumor cells, but causes the tumor cell to act as a factory to produce new virus. These virus 
particles are released as the tumor cell dies, and can then proceed to infect other tumor cells in the vicinity, and 
continue the process of tumor kill. In addition to this direct oncolytic activity, the virus carries a therapeutic 
payload--acting as a gene therapy vector, too--and causes the tumor cell to synthesize and secrete an immunity-
stimulating protein called Interleukin-12 (IL-12) before it is killed. This IL-12 is released and promotes an 
immune response against surviving tumor cells, which increases the antitumor effect of the therapy. The IL-12 
that is expressed can also produce an anti-angiogenic effect, by interfering with the production of new tumor 
blood vessels necessary to allow tumor growth. Anti-angiogenic therapies potentially starve the tumor of 
necessary oxygen and nutrients. Thus, the M032 oHSV produces three different potential mechanisms for 
antitumor effects. The virus has also been genetically engineered to minimize the production of any toxic effects 
for the patient receiving the therapy. 
1.2 Primary Objective 
To determine, in patients who would not be eligible for resection of recurrent glioma, the safety and 
tolerability of stereotactic intracerebral injections of escalating doses of M032 virus (NSC 733972), and to 
determine the maximally tolerated dose (MTD) of M032.  
1.3 Secondary Objectives 
To obtain preliminary information about the potential benefit of M032 in the treatment of patients with 
recurrent malignant gliomas (progression-free survival, overall survival). 
1.4  Tertiary Objectives 
 
1.4.1 
To delineate the local and systemic immune response to M032 administration. 
1.4.2 
To characterize the in situ biologic activity of M032 after intratumoral inoculation, where possible. 
 
2. BACKGROUND 
2.1 Safety and efficacy of non-virulent, oncolytic Δγ134.5 HSV  
The efficacy of herpes simplex virus (HSV) as a treatment for brain tumors has been demonstrated 
experimentally. The earliest studies used a thymidine kinase (tk) deficient HSV vector following the 
demonstration that tk deficient viruses grow relatively slowly in a variety of cell lines, and exhibit reduced 
pathogenicity in situ. Martuza and colleagues demonstrated that subcutaneous and intracerebral tumors regressed 
following treatment with varying titers of a tk- HSV [1-2]. This HSV vector, which consisted of a 360 base pair 
(bp) deletion within the tk gene (resulting in inactivation of the gene), was able to reduce the growth rate of a 
variety of human tumors including gliomas in murine models.  
Additional modified viruses based on the HSV backbone have been developed and tested with encouraging 
results [3-6]. Viruses containing deletions in viral DNA polymerase, the neurovirulence gene (134.5), or HSV-
1/HSV-2 intertypic recombinants also retained the capability of killing cultured human tumors [7].  Specifically, 
the deletion of the neurovirulence gene produced a greater survival of nude mice with human gliomas and retained 
the tk gene, making it susceptible to the antiviral drug acyclovir which is routinely used to treat HSV infections 
[8]. 
  
A series of studies was carried out to investigate variations in the mutation of the neurovirulence gene 134.5.  
Two viruses derived from HSV strain F have been tested exhaustively [5, 9-10].  The first, R3616, has a 1 kb 
deletion in each copy of the 134.5 gene, whereas R4009 has a stop codon inserted in the coding region.  Using a 
severe combined immunodeficiency (SCID) mouse model, Whitley and colleagues demonstrated that both 
viruses, designated Δγ134.5 HSV, were effective agents for the treatment of human gliomas introduced 
intracerebrally into mice.  Moreover, injection of up to 107 plaque-forming units (pfu) of R3616 virus 

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University of Alabama at Birmingham 
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intracerebrally into immunocompetent mice resulted in no deleterious effects, whereas 100 pfu of the parental 
HSV-1(F) virus was lethal.  Additionally, a mutation in a more neurovirulent herpes HSV strain (strain 17) has 
been evaluated extensively [11-12].  Consisting of a 760-bp deletion in both copies of the 134.5 gene, the mutant 
HSV 1716 has also been shown to be aneurovirulent in immunocompetent mice at 106 pfu and to cause regression 
of human brain tumors in mice with corresponding increased survival. 
Martuza and colleagues generated G207, a modified HSV that contains 1) deletions of both copies of 134.5 
and 2) a viral ribonucleotide reductase disabled secondary to disruption of the UL39 gene by insertion of the E. 
coli LacZ coding region [6, 13].  G207 significantly prolonged survival of nude mice bearing human tumors 
either subcutaneously or intracranially when injected at titers of 106-107 pfu. In addition, virus injected 
intracerebrally into HSV sensitive Aotus simian primates did not produce any deleterious side effects, whereas 
<103 pfu of HSV-1(F) virus was lethal. 
Clinical Trials Utilizing Oncolytic Δγ134.5 HSV in Human Malignant Glioma 
G207 and 1716 have both been used in human trials. A dose-escalating phase 1 study of G207 was completed 
in patients with recurrent, progressive malignant glioma [14]. The trial was conducted at University of Alabama 
at Birmingham and Georgetown University Medical Center. Twenty-one patients were enrolled in a total of seven 
dose-escalating cohorts, with three patients per cohort. Patients were stereotactically inoculated with G207 in the 
enhancing portions of their tumors. Five separate loci were inoculated in the final cohort; all previous cohorts 
were inoculated in a single locus within the enhancing tissue only. No toxicity definitively related to G207 was 
observed at doses up to 3 x 109 plaque forming units (pfu). In fact, a toxic dose level was not attained during this 
trial. This was due to viral processing techniques available at that time, limiting the total dose that could be 
administered. 
In a Phase IB study, six patients with recurrent, resectable malignant glioma were enrolled at the University 
of Alabama at Birmingham, in a trial examining a split dose administration strategy of G207 [15]. Patients 
underwent stereotactic inoculation of G207 into their tumors, followed two to five days later by resection of the 
tumor and reinoculation of G207 into the tumor cavity. 1 x 108 pfu was inoculated initially, followed by a second 
dose of 1 x 109 pfu. No dose limiting toxicities were seen in the trial, although one patient suffered a twelve hour 
period of mental status changes, contralateral paresis, and an elevated temperature when a protocol deviation 
resulted in an inadvertent partial dose of virus being administered intraventricularly. The patient fully recovered 
quickly. One of the six patients went nearly two years before remote recurrence of her glioblastoma multiforme 
resulted in her death.   
A third study in 9 patients with recurrent malignant glioma was recently completed in which G207 was being 
administered followed by a single dose of 5Gy of radiation. Nine patients were followed without any dose limiting 
toxicity. Results of this trial are pending completion of a manuscript for submission for publication.   
In a phase 1 study utilizing a similar oncolytic HSV vector, nine patients with relapsed malignant glioma 
received intratumoral inoculation of 1716.  Although 1716 carries only a deletion of the 134.5 gene (it does not 
carry the additional mutation of the UL39 gene found in G207), it also caused no dose-limiting toxicity at all 
doses tested (maximum dose,105 pfu) [16].  Both the G207 and 1716 studies used modified HSV strains  
that retained the tk gene, thus allowing potential response to standard anti-viral therapy if encephalitis occurred. 
Two additional studies of 1716 have been published, neither of which demonstrated any dose limiting toxicities 
[17-18].  
M002 – a murine interleukin-12 expressing Δγ134.5 HSV  
Parker and colleagues have generated M002 from the parent virus R3659, which is similar to both G207 and 
1716 [19]; Figure 1.  R3659 resembles G207 and 1716 except that it carries a 501-bp deletion within the wild-
type tk gene and has, therefore, been constructed with a copy of the HSV-1 thymidine kinase gene at the locus of 
each deleted 134.5 gene.  Additionally, although R3659 is derived from the same wild type parent  
strain as G207, R3659 more closely resembles 1716 in that it does not carry the additional mutation of G207 in 
the UL39 gene. 

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University of Alabama at Birmingham 
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M002 contains deletions of both copies of 134.5 and an insertion in the 134.5 locus of the gene encoding 
murine IL-12 (mIL-12) [20].  The two cDNA sequences encoding the p35 and p40 chains of mIL-12 are co-
expressed by the Egr-1 promoter and linked by an IRES sequence.  Since γ134.5 is a diploid gene, two copies of 
the mIL-12 cassette are present in the virus.  Because the insertion of the mIL-12 gene left the modified R3659 
virus tk-, the deletion in the native tk gene locus was repaired.  For this reason M002 remains sensitive to 
acyclovir. 
M002 has been extensively evaluated in vivo with animal models [20-22].  mIL-12 has been shown to be 
expressed in physiologically relevant concentrations in various tumors of neuroectodermal origin (from 800-3200 
picograms per ml per 24 hours per 5 x 105 cells).  When compared to either the parental virus R3659 which 
expresses no foreign genes or the clinically evaluated HSV G207, M002 produces increased survival in murine 
intracranial tumor models (Figure 2) and increased tumor growth inhibition in subcutaneous models (Figure 3).  
Figure 2:  Immunocompetent C57BL/6xDBA/2 F1 hybrid mice were injected intracranially with 4C8 mouse 
glioma cells, followed in 7 days by saline or the 
viruses shown. Median survivals were determined 
by Kaplan-Meier plots and are shown on the 
Figure.  Log rank analyses of these survival 
values confirmed the significantly prolonged 
survival afforded mice treated with M002, 
compared with R3659 (p = 0.00007) or G207 (p = 
0.0003). In addition, about 20% of the M002-
treated mice were apparently “cured” with no 
evidence of remaining 4C8 glioma cells 
histologically when the experiment was 
terminated.   
 
 
 
 
 
 
 
Figure 2 
 
Days Post Tumor Induction
0
14
28
42
56
70
84
98
112
126
140
154
Proportion Surviving
0.0
0.2
0.4
0.6
0.8
1.0
Saline, 10L
MST = 54 days
G207, 1 x107 pfu/10L
MST = 82 days
M002, 1 x 107 pfu/10L
MST = 97 days
R3659, 1x107 pfu/10L
MST = 79 days
5 x 105 4C8 glioma cells injected intracerebrally
Survivors killed 
at 145 days
(tumor-free)
HSV or Saline
Injected on 
Day 7
 
Figure 1:  Schema #1 is a linear representation of the genomic 
structure of wild-type Strain F HSV-1 from which the other viruses 
were constructed.  Schema #2 represents an intermediate construct 
in which the wild-type thymidine kinase gene is deleted from its 
normal position (UL23 – up arrow in #1) and used to replace the 
diploid gene, 134.5, in the inverted repeat segments.  Schema #3 
depicts the murine IL12 gene construct used to replace the 
heterotopically tk gene.  The IL12 construct is described in the 
adjacent text.  M001 represents the isolate prior to replacing the tk 
gene in its orthotopic location in the ULsegment (M002).   
alan
b
UL
am'c'
b'
US
as
c
a
Ncol BstEll
Ncol
Stul
Ncol
Stul
BspEl
Ncol
BstEll
Ncol
Ncol
p40
IRES p35
Egr-1p
mIL-12 (2.9 kb)
PolyA
TK (1.9 kb)
2287
27p Ncol
R3659
M001/M002
511
1
2
3
a

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 10 of 66 
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Figure 3:  Immunocompetent BALB/c 
mice were injected subcutaneously with 1 
x 105 Neuro-2A cells.  When tumors 
attained 200mm3 in volume, they were 
injected with 50 l of saline or 5 x 107 pfu 
of HSV R3659 or M002.  Tumor volumes 
were measured every 3 days and the 
average percentage increase in volume 
was calculated relative to the treatment 
date.   
 
 
 
 
 
 
 
Preclinical studies have documented 
the increase in efficacy of IL-12 expressing oncolytic HSV in a syngeneic intracranial model of neuroblastoma 
and a second model of syngeneic intracranial murine malignant glioma when compared to a parent γ134.5 deleted 
virus not expressing IL-12 [21]; further studies in both immunocompetent mice bearing syngeneic 
neuroblastomas and nude mice bearing human U87 malignant glioma xenografts have demonstrated the 
superiority of IL-12 expressing oncolytic HSV specifically compared to G207, the virus previously used in 
clinical trials.   Immunohistochemistry shows increased inflammatory response, particularly CD4+ and CD8+ 
cells and macrophages, suggesting that an anti-tumor immune response may be partially responsible for the 
increase   [21]. 
Although M002 has not been used in human clinical studies, the safety of M002 has been addressed both by 
the deliberate molecular engineering of the virus as described above as well as in vivo testing in HSV-sensitive 
owl monkey, Aotus nancymai.  Murine IL-12 has been demonstrated to be active on Aotus lymphocytes, 
validating this as an appropriate model for preclinical safety testing (JM Markert, unpublished data). A purified 
extract (made according to GMP specifications) of M002 has been injected intracerebrally into three Aotus 
nancymai at titers up to 4.8 x 108 pfu with no adverse effects. One of these Aotus is alive and well more than 5.5 
years after inoculation. A second animal was alive and well more than 5 years after inoculation when a positive 
routine TB skin test resulted in the intentional sacrifice of this and another untreated animal in the same colony; 
necropsy later revealed no evidence of tuberculosis infection or viral toxicity.  In contrast, 103 pfu of HSV-1(F) 
virus is lethal.  MRI analysis of M002 treated monkeys showed no CNS abnormalities. Neuropathologic 
evaluation is pending and will be performed at one year post-inoculation. Two additional Aotus have also been 
injected with lower doses of M002 (non-purified standard laboratory purifications of 1 x 107 and 1 x 108 pfu, 
respectively). One of these animals died from iatrogenic antibiotic-induced renal failure and the second from 
anesthetic complications; brains from both animals were fixed and serially sectioned, then stained. No evidence 
of encephalitis or HSV-related toxicity was found. 
M032 – the human Interleukin-12 expressing Δγ134.5 HSV  
Typically, murine proteins have not been used in human gene therapy but mIL-12 retains bioactivity in human 
cells. Unfortunately, human IL-12 has no activity in murine systems, which prevented the use of this protein in 
preclinical (murine) brain tumor models. Reconstruction of the vector to express hIL-12 instead of murine IL-12 
was recommended by the reviewers of our RAID application. This new vector was constructed (Figure 4) and 
extensively characterized, and shown to be similar to M002 in terms of IL-12 production, replication, and 
neurotoxicity in murine models  (JM Markert, GY Gillespie, unpublished data)  [20, 22].  Moreover we have 
determined that the recombinant human IL-12 produced by M032 HSV-infected cells is biologically active and 
able to stimulate the proliferation of primed Aotus peripheral blood lymphocytes as evidenced by tritated 
thymidine uptake [41].  
 
 
Days After Virus Injection
0
2
4
6
8
10
12
14
16
18
2
Average Percentage Size
 Increase
0
1000
2000
Saline
R3659
M002

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
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As M032 represents an excellent vector for evaluation in clinical studies for the treatment of glioblastoma 
multiforme, the NIH Rapid Access to Investigational Drug (RAID) Program agreed to produce a M032 Master 
Viral Bank, manufacture a toxicology lot of the virus, conduct biodistribution and toxicology studies in the HSV-
hypersensitive non-human primate, Aotus nancymae, and to produce and qualify a clinical lot of virus in 
accordance with Good Manufacturing Practice (GMP) regulations and tested according to the Points to Consider 
for the Testing of Cell Lines Used in the Manufacture of Biological Products [23].  These and other documents, 
including this protocol, were used to obtain an IND from the FDA, #14946.  
The Biodistribution and Toxicology studies were conducted by intracerebral injection of 100 microL of M032 
HSV into 30 of 36 Aotus nancymae monkeys, distributed evenly by gender.  The remaining 6 monkeys were 
injected with saline as a control.  Two doses of HSV were tested – a low dose of 1 x 106 plaque-forming units 
(PFU) and a high dose of 1 x 108 PFU.  On a dose/gram of brain basis, these doses would correspond to human 
equivalent doses of 5 x 107 PFU and 5 x 109  PFU.  Two male and two female monkeys in each of the low dose 
and high dose groups were killed at 3, 31 and 91 days and a complete necropsy was performed, including PCR 
analysis for presence and copy number of HSV in various organs and body sites.  There were no serious adverse 
events or adverse events that could be attributed directly to the test article, M032.   
However, one animal was killed at 16 days due to a progressively moribund condition.  This animal was 
losing weight, not eating and progressively lethargic, without clear clinical evidence of viral encephalitis (no 
fever).  A complete necropsy did not reveal any histopathological changes indicative of HSV encephalitis and 
there were no substantially elevated levels of HSV as determined by quantitative PCR in the brain or in other 
organs, compared to the levels detected in all other monkeys.  While there was nothing to support a diagnosis of 
HSV encephalitis or HSV infection in other organs of the body, nevertheless, neither could it be excluded that 
the test article was not a contributing cause of this monkey's deteriorating condition 16 days after the injection.  
In summary, the HSV M032 that will be used in this clinical trial has been qualified for safety and stability and 
has been determined to be safe for intracerebral administration in HSV-hypersensitive non-human primates at a 
dose that far exceeds that we intend to employ here.     
2.2 Malignant Glioma 
Malignant gliomas are the most common primary brain tumors of humans, accounting for 30% of all primary 
central nervous system (CNS) tumors in adults.  There are three types of malignant gliomas: (i) anaplastic 
astrocytoma, (ii) glioblastoma multiforme and (iii) gliosarcoma. Primary malignant brain tumors in the United 
States are estimated to occur at an incidence of 14.7 per 100,000 people, and 10,000—15,000 new  
cases are diagnosed annually [24]. Glioblastoma multiforme (GBM), the most malignant type of brain tumor, has 
been refractory to improvements in treatment; the outcome of conventional treatments is poor, with a median 
survival of one year. 
 
Gliomas develop from the unsuppressed growth of neural support cells called astrocytes, usually resulting 
in the first clinical symptoms such as memory loss, visual impairment, or seizures. Presently, the most common 
modalities for the treatment of brain tumors are surgical resection, chemotherapy and/or radiation therapy, 
depending on the severity of the tumor and type of malignancy. The major problem associated with conventional 
treatment stems from tumor recurrence. 
 
FIGURE 4  
 
 
 
Figure 4:  The 134.5 gene is diploid, one 
copy in each of two inverted repeat segments.  
Construction of M032 was achieved by 
replacing both copies of the 134.5 gene by 
homologous recombination using flanking 
sequences cloned into the 5’ and 3’ ends of 
human IL-12 construct described above.  
Terminal repeat insertion only shown for 
simplicity. The M032 HSV construct was 
repaired to express native HSV thymidine 
kinase at the UL24 position (Up arrow); the 
virus is sensitive to anti-viral drugs such as 
acyclovir or valgancyclovir. 

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University of Alabama at Birmingham 
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2.3 Rationale 
Many vectors have been considered for neoplastic therapy, including naked DNA, liposomes, and viruses. 
Viral vectors include those derived from retrovirus, adenovirus (AdV), adeno-associated virus (AAV) and HSV 
type 1. Each of the vectors has advantages and disadvantages for treating glioma in situ. 
Recombinant HSV have been generated containing a variety of gene deletions. Their effectiveness as gene 
therapeutic agents have been examined in vitro and in vivo in numerous models of metastatic and primary tumors. 
These experiments have provided valuable safety information in various animal models. HSV vectors are 
neurotropic, for they have evolved to coexist within neuronal tissue (latency).  The identification of genes 
essential for replication in the brain [8, 25] has opened new therapeutic avenues for currently incurable brain 
tumors.  It has been clearly demonstrated experimentally that by altering these genes, either by deletion, insertion, 
or point mutation in the coding region, the virus will kill dividing tumor cells but is otherwise avirulent following 
delivery into post-mitotic brain tissue.  Thus, it is possible to develop a genetically stable engineered virus with 
the ability to selectively replicate in glioma cells.  This property is desirable when considering that, following 
resection, more than 109 tumor cells may remain in situ.  Other methods of treatment, e.g., a retroviral vector 
with the HSV tk gene, have been unable to target this many cells, especially in light of the ongoing cellular 
proliferation of the remaining glioma cells.  However, the ability of the modified HSV to selectively replicate 
within the tumor may allow for the delivery of sufficient virus into the tumor bed to potentially destroy tumor 
cells remaining after conventional therapy. 
As described in Section 2.1, one conditionally replicating HSV, G207, has been tested by intracerebral 
administration in humans at doses up to 3 x 109 pfu without evidence of toxicity that could be attributed to the 
vector.  However, because the selectively replicating recombinant HSV can still carry other recombinant genes, 
the opportunity exists to combine its direct oncolytic effects with additional gene therapy effects.  The approach 
utilized with this study agent, M032, depends upon the modified virus’ direct oncolytic activity in combination 
with the recruitment of a targeted inflammatory response to kill the malignant tumor cells. 
This “double-barrel” effect is accomplished by the expression of IL-12 in physiologically relevant 
concentrations from cells infected with the recombinant HSV [20-22].  Murine IL-12 is known to cross-react 
with the human IL-12 receptor and stimulate an immune response, but human IL-12 does not cross react with the 
murine receptor; thus all in vivo preclinical efficacy studies have necessarily been performed in murine models 
using M002, a variant of the virus that expresses murine IL-12 [20-22]. IL-12 is a cytokine with potent antitumor 
qualities [26-28].  By induction of interferon- production [29], IL-12 acts to enhance the cytolytic activity of 
natural killer cells and cytotoxic T lymphocytes [30].  IL-12 also induces a TH-1 type immune response [31], 
which may provide a more durable antitumor effect than other cytokine anti-neoplastic approaches.  Furthermore, 
IL-12 has also been demonstrated to have in vivo anti-angiogenic activity which may contribute to its antitumor 
effects [32-33].  It has been shown to produce important antiglioma activity in different rodent models [34-35].  
In preclinical models, M002 has been shown to be more effective than G207 or similar viruses as an inhibitor of 
tumor growth.  Immunohistochemistry studies suggest that this increased efficacy may be due to an anti-tumor 
immune response induced by the secretion of IL-12 by the infected tumor cells [36]. A potential advantage of IL-
12 is its ability to stimulate an antitumor response in the absence of IL-2. This is important in patients with 
malignant glioma, who suffer from a decreased ability to respond to IL-2. Nonetheless, the local 
immunomodulatory effects of IL-12 in the immunosuppressive milieu of human malignant gliomas is currently 
unknown, and will only be understood through a phase 1 study designed to assess biologic activity. 
 
While early clinical studies of systemic IL-12 have produced evidence of toxicity [35], its effects when 
administered locally are not well known.  As discussed in Section 2.1, preclinical models support the safety of 
intracranial inoculation with M032 in two relevant species (mouse, non-human primate). 
 
3. PATIENT SELECTION 
3.1 Eligibility Criteria 
3.1.1 
Patients must have histologically or cytologically confirmed glioblastoma multiforme, anaplastic 
astrocytoma, or gliosarcoma. 

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3.1.2 
Prior therapy:  Patients must have failed external beam radiotherapy to the brain, and if eligible and 
tolerated, undergone appropriate treatment with temozolomide chemotherapy. All radiation and 
additional chemotherapies must have been completed at least 4 weeks prior to enrollment. Prior 
therapy with nitrosoureas must have been completed at least 6 weeks prior to enrollment.  
3.1.3 
Age ≥18 years (age of majority for clinical trials in Alabama).  Because no dosing or adverse event 
data are currently available on the use of M032 in patients <18 years of age, children are excluded 
from this study but will be eligible for future pediatric phase 1 single-agent trials. 
3.1.4 
Karnofsky Performance Status (KPS) ≥70% (see Appendix B). 
3.1.5 
Life expectancy of greater than 4 weeks. 
3.1.6 
Patients must have normal organ and marrow function as defined below: 
 
leukocytes……………………......>3,000/μl 
 
absolute neutrophil count ........... ..>1,500/μl 
 
platelets ...................................... ..>100,000/μl 
 
total bilirubin .............................. ..within normal institutional limits 
 
AST(SGOT)/ALT(SGPT)….. .... ..<2.5 X institutional upper limit of normal 
 
creatinine within normal institutional limits 
 
 
OR 
creatinine clearance……………..>60 mL/min/1.73 m2 for patients with creatinine levels above 
 
  institutional normal. 
3.1.7 
Residual lesion must be ≥1.0 cm and < 5.5 cm in diameter without bilateral extension through the 
corpus callosum as determined by MRI as this is a locally delivered treatment. These parameters will 
be re-evaluated on imaging done on the day of catheter implantation and if the lesion no longer meets 
the criteria, the patient will not undergo catheter implantation or treatment with M032. 
3.1.8 
The effects of M032 on the developing human fetus are unknown.  For this reason, women of child-
bearing potential and men must agree to use adequate contraception prior to study entry and for the 
first six months after receiving M032.  Because it is currently unknown if M032 can be transmitted 
by sexual contact, a barrier method of birth control must be employed and for six (6) months 
following the administration of the study drug.  Should a woman become pregnant while participating 
in this study, she should inform her treating physician immediately.  For two weeks after receiving 
M032, subjects should avoid intimate contact with pregnant women, infants and young children and 
individuals with decreased immunity (ability to fight infection). Subjects should also refrain from 
donating blood during the trial. 
3.1.9 
Ability to understand and the willingness to sign a written informed consent document. 
3.1.10 Females of childbearing potential must not be pregnant; this will be confirmed by a negative serum 
pregnancy test within 14 days prior to starting study treatment. 
3.1.11 Steroid use is allowed as long as dose has not increased within 2 weeks of scheduled M032 
administration.  Whenever possible, the patient should be on a steroid dose that is equivalent to a 
dexamethasone dose of ≤ 2mg daily at the time of treatment. 
3.2 Exclusion Criteria 
3.2.1 
Patients who have had chemotherapy, cytotoxic therapy, immunotherapy within 4 weeks prior to 
entering the study (6 weeks for nitrosoureas), surgical resection within 4 weeks prior to entering 
the study, or have received experimental viral therapy or gene therapy at any time (e.g., adenovirus, 
retrovirus or herpes virus protocol).  However, this does not preclude re-treatment with M032 at a 
later date.  
3.2.2 
Patients who have not recovered from adverse events due to therapeutic interventions administered 
more than 4 weeks earlier. 
3.2.3 
Patients may not be receiving any other investigational agents. 

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M032 Amendment 12 
University of Alabama at Birmingham 
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3.2.4 
History of allergic reactions attributed to compounds of similar biologic composition to M032 or to 
IL-12. 
3.2.5 
Tumor involvement which would require ventricular, brainstem, basal ganglia, or posterior fossa 
inoculation or would require access through a ventricle in order to deliver treatment. Also, since 
M032 is a local treatment, patients whose tumors have bilateral extension through the corpus 
callosum, those with actively growing multifocal disease by MRI, and/or CSF dissemination/ 
leptomeningeal disease, are ineligible, .  
3.2.6 
Prior history of encephalitis, multiple sclerosis, or other CNS infection. 
3.2.7 
Required steroid increase within 2 weeks of scheduled M032 administration. When possible, the 
patient should be on a dexamethasone equivalent dose of ≤ 2mg daily at the time of treatment. 
3.2.8 
Active herpes lesion. 
3.2.9 
Concurrent therapy with any drug active against HSV (acyclovir, valacyclovir, penciclovir, 
famcyclovir, gancyclovir, foscarnet, cidofovir). 
3.2.10 Uncontrolled intercurrent illness including, but not limited to ongoing or active infection, 
symptomatic congestive heart failure, unstable angina pectoris, cardiac arrhythmia, or any other 
medical condition that precludes surgery.  Also, psychiatric illness/social situations that would limit 
compliance with study requirements. 
3.2.11 Excluded patient groups: 
Pregnant women are excluded from this study because M032 is a viral oncolytic therapy with 
unknown potential for teratogenic or abortifacient effects.  Because there is an unknown but potential 
risk for adverse events in nursing infants secondary to treatment of the mother with M032, 
breastfeeding women will not be included in the study. 
Because patients with immune deficiency will be unable to mount the anticipated immune response 
underlying this therapeutic rationale, HIV-seropositive patients are excluded from this study.  
Other treatment studies for this disease that are less dependent on the patients’ immune response 
are more appropriate for HIV-seropositive patients. 
3.2.12 Patients with known history of allergic reaction to IV contrast material that is not amenable to pre-
treatment by UAB protocol. 
3.2.13 Patients with pacemakers, ferro-magnetic aneurysm clips, metal infusion pumps, metal or shrapnel 
fragments or certain types of stents. 
3.2.14  Receipt of Gliadel Therapy.  
3.2.15 Receipt of Bevacizumab (Avastin) therapy within 4 weeks of scheduled M032 administration. 
(Receipt of Bevacizumab (Avastin) greater than 4 weeks of scheduled M032 administration does not 
exclude patient. 
3.2.16  Any other reason the investigator deems subject is unfit for participation in the study 
3.3 Inclusion of Women and Minorities 
Both men and women and members of all ethnic groups are eligible for this trial.  The proposed study 
population, assuming the maximum number of patient enrollments occurs, is illustrated in the table below. 
Gender 
Asian or Pacific 
Islander American 
Indian or Alaskan 
Native 
*Black, not of 
Hispanic Origin 
 
Hispanic 
White, not of 
Hispanic 
Origin 
Other or 
Unknown 
(Non-white) 
 
*Total 
Female 
-- 
1 
1 
8 
-- 
10 
Male 
-- 
1 
0 
9 
-- 
10 
TOTAL 
-- 
2 
1 
17 
-- 
20 

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Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
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*Based on percentages of patients with recurrent malignant gliomas treated at UAB 2000-2015 
4. TREATMENT PLAN 
4.1 Study Summary 
This will be an open-label, CRM-directed, dose-escalating Phase I safety trial conducted at a single site 
(UAB) in patients with a confirmed diagnosis of progressing MG after TMZ chemoradiation therapy. Treatment 
will be administered on an inpatient basis.  Expected adverse events caused by M032 are described in Section 5.  
No investigational or commercial agents or therapies other than those described below may be administered with 
the intent to treat the patient’s malignancy. 
These subjects will include eligible patients for whom surgical resection of their recurrent glioma is 
considered to be infeasible or would not likely afford greater benefit than the potential risks associated with 
surgical resection.  All patients will be required to be placed on an anticonvulsant at the time of screening; those 
patients already on an anti-convulsant (anti-epileptic drug/AED) will have a second anti-convulsant added per 
the discretion of the investigator. This anti-convulsant will be continued for a minimum of 28 days unless the 
patient develops a reaction to the medication. If this develops, all attempts will be made to implement an 
alternative agent. On Day 0, these subjects will be treated under monitored local anesthesia, or at the surgeon’s 
discretion, under general anesthesia. The patients will undergo a contrasted MRI scan to determine the site(s) for 
stereotactic biopsy and inoculation of the test agent.  Patients will then undergo stereotactic biopsy of their tumor.  
While evidence of radiation damage or necrosis may be present on the frozen section, inoculation with M032 
will only proceed if viable, recurrent glioma is also present on the frozen section. 
These patients will undergo placement of one to four stereotactically placed catheters, which will be primed 
in the OR with 0.4 ml preservative-free normal sterile saline solution per catheter.  The wound will be closed, the 
catheters aseptically exteriorized and the subject allowed to recover overnight from the procedure in the 
Neurosurgery ICU.  If the patient’s condition is stable they may be transferred the next morning, Day 1, to a 
Clinical Research Unit bed for M032 administration and monitoring until discharge.  After pathology results 
confirm recurrent tumor, and a CT confirms appropriateness of catheter placement, infusion of M032 may begin. 
If a catheter is determined, by the investigator, to be out-of-position and cannot be properly positioned by simple 
partial withdrawal at the bedside (as determined by the surgeon investigator), this/these catheter(s) will not be 
used for infusion.  The catheters will be removed 6-18 hours after the completion of the infusion. 
Patients will then receive the entire M032 dose by intratumoral infusion over a 6 hour period (infusion may 
extend up to 12 hours, if needed due to catheter malfunction).  The total amount of M032, as defined by each 
patient’s dose level, will be delivered in a total volume of ~2.4 ml administered in up to four catheters, each 
attached to a separate syringe mounted in an approved syringe pump for a total infusion rate of 400 microliters 
(0.4 ml) per hour. However, the infusion of the first syringe(s) will actually last 3 hours and 35 minutes to allow 
flushing of the saline solution from the catheter(s).  The infusion rate for the flush will be 400 microliters (0.4 
ml) per hour for 35 minutes per catheter. After the 35 minute flush, the infusion rate will be changed to the 
appropriate rate for the M032 infusion. Subsequent to this, the rate of administration through each individual 
catheter shall be determined by the equation: 
400 microliters (0.4 ml) per hour/number of active catheters. 
 Each catheter will be placed in a different enhancing area of the tumor and labelled.  If, in the surgeon’s 
judgment, the tumor location or other properties are not suitable for all four catheters to be placed, fewer 
catheters will be placed, ranging from number one to four.  
 
The total volume administered from each syringe will be determined by the number of catheters actively 
infusing, using a volume of 400 microliters (0.4 ml) per hour as a guide.  Each initial loaded syringe will be 
replaced at 3 hours 35 minutes after the infusion is started with a new loaded syringe that has been maintained at 
4oC and new infusion tubing.  Refer to table below. 
Table :  Plan for Loading rate and Infusion Rate for M032 HSV 
Number of 
Catheters 
Initial Loading Rate for First 35 mins per catheter Final Infusion Rate per catheter 
1 
0.4 cc/hour 
0.4cc/hour 

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M032 Amendment 12 
University of Alabama at Birmingham 
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For each of the study drug infusion periods [active catheter(s)/pump(s)], the study drug syringe will be 
connected to the infusion tubing, that has been flushed with sterile PBS + 10% glycerol (Alanza) . The infusion 
tubing will then be primed with 4.6 ml of the diluted M032 virus preparation in the syringe and connected to the 
catheter. The first infusion period will begin with a 35 minute flush of the saline solution from the catheter at a 
rate of 0.4ml/hour/catheter.  After completion of the flush, the pump will be reprogramed to the appropriate rate 
for the study drug infusion and the infusion begun.  After the first period of infusion is complete, the infusion of 
the study drug from a single syringe at 400 microliters/hour should be 1.2ml.  The infusion will be stopped, and 
the above procedure (minus the flush) repeated for the second syringe that has been stored at 4oC and new infusion 
tubing.  After two infusion periods, the delivery of the total planned dose should be complete. 
If delivery at the desired rate is not possible through an individual catheter, the rate of delivery through that 
catheter may be slowed and the rate of delivery through the remaining catheters increased as possible to attempt 
to maintain total delivery rate at 400 microliters (0.4 ml) per hour. 
After the initiation of the infusion, the subjects’ vital signs (temperature, blood pressure, pulse, respiratory 
rate) and neurologic function and limited neurological exam will be monitored every hour for the first twelve 
hours, then every two hours for the next 12 hours, then every 4 hours for the next 24 hours. Thereafter, the 
frequency of monitoring will be determined by the attending physician(s) based on the medical condition of the 
patient.   
After discharge the subject will continue to be closely followed for evidence of adverse events, with 
outpatient follow-up evaluations scheduled at Day 10 and months 1, 2, 3, 4, 5, 6, 9 and 12, or more often if 
medically indicated. HSV detection will be performed at additional time points. The visits at months 2, 4 and 5 
will consist of sample collections and a study coordinator visit only. Due to the complicated procedure for 
obtaining samples remotely at visits 2, 4 and 5, if a patient is unable to travel then, and if determined feasible by 
the PI, an attempt can be made to have the specimens collected by the patients’ referring physician and sent to 
UAB for analysis. 
4.1.1 Continual Reassessment Method 
Dr. Gary R. Cutter and Dr. Inmaculada B. Aban, Professors in the Department of Biostatistics, UAB School 
of Public Health, with expertise in CRM calculations will apply the software developed by Dr. Steven Piantadosi, 
(Cedars-Sinai) who will serve as a clinical trial statistical consultant to advise and assist in the implementation 
of the Continual Reassessment Method in determining dose alterations. In addition, Drs. Aban and Cutter, will 
provide support in clinical trial and data management, and data analysis. 
Rationale for the Use of a Modified Continual Reassessment Method: The description of the determination 
of MTD from the CRM that we are employing is adapted from that used previously in a Phase I/II study of the 
poly (ADP-ribose) polymerase-1 (PARP-1) inhibitor BSI-201 in patients with newly diagnosed malignant glioma 
conducted under the New Approaches to Brain Tumor Therapy (NABTT) CNS Consortium. To estimate the 
MTDs in terms of clinical toxicities, a modified continual reassessment method (CRM), based on that described 
by Piantadosi et al [45], will be employed. The CRM has been shown to be less biased and more efficient for 
estimating the MTD than traditional dose-finding models. The efficiency of the CRM stems from its explicit use 
of biological knowledge in the form of a parametric dose toxicity model. In the CRM, only a starting dose is 
specified and the dose is escalated or deescalated based on the toxicities of the previous cohort. 
This is advantageous because the investigators can choose the next dose level based on all available clinical 
and statistical information rather than on relatively arbitrary predetermined doses. One disadvantage of the CRM 
is uncertainty of the dose levels that will be used in the trials, because the specific dose levels are not defined at 
the start of the trial.  
Details of CRM Design:  The primary statistical outcome for this study is the occurrence of serious clinical 
toxicity. Investigators would like to employ a dose of drug that yields approximately a 1/3 chance of serious 
toxicity. In our opinion, this represents the best chance at a beneficial therapeutic ratio for this unusually difficult 
disease to treat. This target probability of toxicity is chosen based on knowledge of previous trials of oHSV in 
2 
0.4 cc/hour 
0.2cc/hour 
3 
0.4 cc/hour 
0.13 cc/hour 
4 
0.4 cc/hour 
0.1cc/hour 

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glioma, but is fundamentally subjective. The relationship between dose of drug and probability of toxicity is 
assumed to follow a two-parameter logistic model, both before and during the dose finding given by:  
   
where β is the “slope” of the curve and d50 the midpoint (or dose that yields a 50% response). The parameters β 
and d50 govern the dose finding process, and are estimated, denoted by 𝛽̂ and 𝑑̂50, initially by clinical judgment 
and afterwards by the observed data. This represents a second subjective component of dose finding designs.  
Classical dose-ranging designs incorporate this component of subjectivity by setting out, in advance of the 
experiment, a set of doses to employ. A strictly Bayesian approach to the CRM requires a joint prior probability 
distribution for the parameters β and d50. However, we have found it 
easier for clinicians to render their clinical judgment about β and d50 in 
the form of pseudo-data. Specifically, we ask for two points on the dose 
toxicity curve: the dose thought to yield a 10% probability of toxicity (d10) 
and the dose thought to yield a 90% probability of toxicity (d90). Two 
points, d10, d90, are required to initiate the fit for a two-parameter model. 
The exact points chosen are arbitrary, but we have found these generally 
work well or are easily adapted. The d10 point can often be discarded after 
some data have been obtained. The d90 point is necessary to obtain a fit of 
the model, but is usually relatively unimportant because it characterizes a 
region of the model that does not heavily influence prediction of the next 
dose. In this sense, d90 is a nuisance parameter – necessary for model 
fitting, but relatively uninfluential on the dose escalation.  
At both d10 and d90, the investigator specifies a numerator (number of 
responses) and denominator (number of subjects treated), such that 
numerator/denominator = 0.1 or 0.9 as the case may be. The denominators 
are always taken to be small numbers, e.g. 1, to represent weak evidence 
and reduce the influence of these points compared to real subject data. 
Three values for d90 will be chosen in this protocol with weights of one tenth that of real subject data.  This allows 
flexibility in the upper end of the dose toxicity curve. This adaptation to the CRM came about after subject data 
demonstrated that d90 was incorrect in previous studies, because doses approaching d90 were tried with no 
toxicities observed. This situation required that d90 be moved to a higher dose or the fitted dose toxicity curve 
became too steep and no dose escalations resulted.  
We assume the dose for d10 = 1 x 104 pfu for M032 (Table 1). A set of three values will be chosen for d90 =1 
x 1015 pfu. These values for d90 spread probability mass over a wider range and may eliminate the need to increase 
d90 if the data are inconsistent with a single value. Based on these starting points, our desire is to employ a dose 
of virus to yield no more than 33% chance of clinical toxicity. The initial starting dose will be assumed to be 1 x 
105 PFU and the maximum dose to be tested would be 1 x 109 PFU. Further assumptions would be that the 
maximum number of subjects would be 204 and that the dose cohort size would be 1.  Further, the next CRM 
recommended log dose would be rounded to the nearest 0.5 or full dose. Moreover, the number of consecutive 
subjects at the maximum tolerated dose would be 10 or a number until 20 subjects are treated, whichever comes 
first.  This will allow us to collect more information of the safety of the MTD.  If a second toxicity is observed 
for patients on the highest dose, then the next lower dose is used and declared MTD if 10 consecutive subjects 
on the same dose are toxicity free or if n=20 is reached. If more toxicities are found, CRM will be used in the de-
escalation of the dose. 
The outcome space for the first CRM iterations would look like the values shown as six sample scenarios in 
Table 2 using a starting dose 1 x 105 pfu. We will not allow dose escalations to exceed 10 times the maximum 
dose already administered. In this way, we prevent a too rapid dose escalation or testing high doses without a 
reasonable degree of certainty concerning their safety. Although the values in Table 2 appear reasonable to us 
currently, we emphasize that they should not be taken literally. Indeed, if we receive approvals (DSMB, IRB, 
FDA), we will update the CRM model using the four patients previously enrolled and treated.  Some of the data 
Table 1: Initializing Data used 
to start the CRM  
Log 
Dose 
N 
r 
Weight 
4 
1 
0.05 
0.5 
5 
1 
0.10 
0.5 
6 
1 
0.2 
0.5 
7.5 
1 
0.5 
0.5 
9 
1 
0.8 
0.5 
10 
1 
0.9 
0.5 
12 
1 
1 
0.5 
15 
1 
1 
0.5 
(Initial recommended dose based on 
these data=6.7842) 

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could suggest that the current value for d10 and/or d90 is inappropriate. We would not proceed blindly with a CRM 
(or any other) dose escalation in such a circumstance.  All testing doses will be the estimated dose round-down 
to the nearest 10th.  
For each subsequent level, the investigators will evaluate the recommended dose-level. It is possible that the 
model may recommend large interval increases in dose level with which the investigators do not feel comfortable. 
In these cases, the investigators will use this information and their best clinical judgment to assign a next dose 
level. We emphasize that the 
dose levels recommended by 
the CRM should not be taken 
literally. 
Model 
based 
dose 
escalation methods, especially 
the CRM, are able to account 
for ordinal or quantitative 
toxicity assessments, and use 
this 
information 
to 
guide 
subsequent dose changes. In 
particular, 
the 
binomial 
likelihood is 
where i indexes the dose 
level, p is the probability of 
toxicity (equation 1 above), d is 
dose, n the number of subjects 
treated at each dose, and r the 
number of toxicities. While n is 
constrained to be an integer, r 
does not have to be integral. In 
particular, r can be taken to be 
the 
sum 
of 
ordinal 
or 
quantitative toxicity measures 
provided that r < n. The 
maximization of equation 2 
with respect to the parameters 
(model 
fitting) 
can 
then 
proceed in the usual fashion. 
Furthermore, the ordinal scores 
need not be the same for all 
types of toxicity. For example, we might not want alopecia to have the same effect on dose reduction as neurologic 
toxicities.  
As subjects are treated at doses estimating the true MTD, the recommended subsequent dose levels will begin 
to converge. The criteria to declare the MTD will be when 2 recommended doses are within 10% of one another. 
It is possible that the dose escalation could continue indefinitely and that the MTD is not reached. If 10 subjects 
are treated in the full dose range in this dose-finding study and the MTD has not been reached, we will take pause, 
evaluate the data in consultation with the DSMB, and determine whether to continue the dose-escalation or 
terminate the Phase I portion of the study. If a MTD is not being reached based on clinical toxicities (too many 
or too few), the biological indicator (increased PFS or OS) may factor in the choice of the M032 dose for the 
ultimate planned phase II trial.  
A maximum of 10 subjects will be treated at the putative MTD to have better estimation of ≤ 33% DLT rate.  
Table 2:  Six Different CRM-defined Dose Escalation Scenarios 
Subject 
# 
S1 
S2 
S3 
S4 
S5 
S6 
1 
5 
5 
5 
5 
5 
5 
2 
6 
6 
6 
6 
6 
6 
3 
7 
7 
7 
7 
7 
7 
4 
7.5 
7.5 
7.5 
7.5 
7.5 
7.5 
5 
8 
8 
8 
8 
8 
8 
6 
8 
8 
8 
8 
8 
8 
7 
8.5 
8.5 
8.5 
8.5 
8.5 
7.5 
8 
9 
9 
9 
9 
8 
8 
9 
9 
8.5 
9 
9 
8.5 
8 
10 
9 
8.5 
9 
9 
8.5 
8 
11 
9 
8.5 
9 
8 
8.5 
8.5 
12 
9 
8.5 
9 
8.5 
9 
8.5 
14 
9 
8.5 
9 
8.5 
9 
9 
15 
9 
8.5 
9 
8.5 
9 
9 
16 
9 
8.5 
9 
8.5 
9 
9 
17 
9 
8.5 
8.5 
8.5 
9 
9 
18 
9 
8.5 
8.5 
8.5 
9 
9 
19 
 
8.5 
8.5 
8.5 
9 
9 
20 
 
 
8.5 
8.5 
9 
9 
MTD 
9 
8.5 
8.5 
8.5 
9 
9 
(yellow cells= maximum tolerated dose (MTD); magenta cells =toxicity; 
white cells=no toxicity) 

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No enrollment of special or vulnerable populations is anticipated. 
4.1.2 
Estimated Number of Patients 
Moving forward, we anticipate as few as four and as many as 20 additional patients could be enrolled and 
treated in this trial. We expect approval of the use of the four already enrolled and treated patients to seed the 
model, which would bring the overall total range from 8- 24 patients. If the highest planned dose is successfully 
administered to subjects and assuming one subject per CRM-calculated dose level (10), and an additional 10 
subjects will be entered at the CRM-estimation of the safe target dose (10+10 = 20).     If all subjects develop 
DLTs, the CRM model would de-escalate both through the initial and the two lower dose levels involving no 
more than 4 subjects and the trial would be halted (Section 4.5) with a minimal accrual of 4 subjects.   
Assuming that no DLTs will be encountered, we expect that under optimal conditions only 20 subjects (10+10) 
will be enrolled and treated in this study. These numbers will be modified if the new patient data for seeding 
the model is approved and could even decrease. 
Thus, the maximum number of subjects that we expect to enroll and treat in this trial would be 20, assuming 
that the CRM will adjust each dose after the first 2 or 3 subjects and as more subjects are enrolled to encompass 
the initial test range.   On the other hand, the minimum number of subjects who could be enrolled in this trial 
would be 4 assuming 2 DLTs at Dose Level 1, and the CRM recommending one or two lower doses.  
4.2 Definition of Dose-Limiting Toxicity 
The Cancer Therapy Evaluation Program (CTEP) has published Common Terminology Criteria for Adverse 
Events, version 4.0 for the grading of adverse events experienced in clinical trials for anti-neoplastic agents.  Any 
grade 3 or greater non hematologic toxicity as defined by NCI CTCAE v 4, and considered as being possibly, 
probably or likely related to M032 will be considered a Dose Limiting Toxicity (DLT). However, specific 
symptoms including seizures, neurologic deficits such as weakness, dysphasia or sensory loss are prominent in 
this patient population due to the fact that these tumors frequently disrupt the function of normal areas of eloquent 
brain. Neurologic symptoms from involvement of eloquent brain are commonplace; corticosteroid, radiation, and 
chemotherapies also produce pre-existing symptoms in almost all cases. The pre-existing conditions listed in the 
Table on the following page are acceptable for inclusion in the clinical trial, provided the subjects meet the other 
inclusion and exclusion criteria. 
For all of these pre-existing conditions, DLTs will be defined as progressive involvement of neurological 
dysfunction not attributable to tumor progression and/or the development of life threatening symptomatology 
associated with the original baseline status.  
Neurologic deterioration will not be considered dose-limiting if it resolves back to the patient’s baseline 
within two weeks of completing treatment. For example, increased weakness or aphasia that occurs during the 
infusion will be treated with increasing steroid doses and/or slowing of infusion rates, but not treated as a DLT 
if symptoms improve to baseline within fourteen days of completing treatment. Because of the site of M032 
inoculation, important additional events that will be considered dose-limiting toxicities if they are likely related 
to M032 include death, stroke, hematoma requiring surgery, untreatable neurologic deterioration, unresponsive 
systemic infection, and disseminated HSV infection. 
Management of the above adverse events is outlined in Section 5. 
The maximally tolerated dose (MTD) is the highest dose level below the maximally administered dose when 
dose escalation decisions are made according to the guidelines above.  Enrollment will be stopped when the MTD 
is determined or the maximally planned dose is acceptably tolerated by six patients. 
There will be a minimum twenty-four (24) day observation period between the first patient at each dose level 
and subsequent patients enrolled to allow for evaluation of potential toxicity. If no dose-limiting toxicity is seen 
with the first patient, the twenty-four day waiting period will be waived for the second and third patients at that 
dose level.  If the incidence of DLT at any given dose level meets the criteria for dose escalation, there will be a 
waiting period of a minimum of 24 days before dose escalation occurs.  The Medical Monitor, in conjunction 
with a duly chartered external Data and Safety Monitoring Board comprised of an Infectious Disease expert, a 
neurosurgeon familiar with oncolytic virus therapy, a neuro-oncologist familiar with oncolytic virus therapy and 
an expert in the use of molecular biology methods to detect and quantify Herpes Simplex Viruses in clinical 

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specimens, will evaluate the safety of each dose tested to determine whether the protocol may proceed to the next 
dose level.  
 
CTCAE# 
Body System 
Event name 
Definition of Grade 3 
Toxicity 
Rationale for inclusion 
222 
General disorders 
and administration 
site conditions 
Gait disturbance 
Disabling; limiting self 
care ADL 
Many subjects with malignant glioma have 
gait disturbance as a result of chronic 
corticosteroid administration 
494 
Musculoskeletal and 
connective tissue 
disorders 
Muscle weakness 
left-sided 
Limiting self care ADL; 
disabling 
Malignant glioma can produce muscle 
weakness due to interruption of motor 
function 
495 
Musculoskeletal and 
connective tissue 
disorders 
Muscle weakness 
lower limb 
Limiting self care ADL; 
disabling 
Malignant glioma can produce muscle 
weakness due to interruption of motor 
function 
496 
Musculoskeletal and 
connective tissue 
disorders 
Muscle weakness 
right-sided 
Limiting self care ADL; 
disabling 
Malignant glioma can produce muscle 
weakness due to interruption of motor 
function 
498 
Musculoskeletal and 
connective tissue 
disorders 
Muscle weakness 
upper limb 
Limiting self care ADL; 
disabling 
Malignant glioma can produce muscle 
weakness due to interruption of motor 
function 
527 
Nervous system 
disorders 
Ataxia 
Severe symptoms; limiting 
self care ADL; mechanical 
assistance indicated 
Malignant glioma itself and many of the 
anti-epileptics drugs used to treat seizures 
associate with this disease can produce 
ataxia that requires mechanical assistance 
529 
Nervous system 
disorders 
Central nervous 
system necrosis 
Severe symptoms; medical 
intervention indicated 
Accepted treatments for malignant glioma 
(for example radiation therapy) can produce 
CNS necrosis that requires medical 
intervention such as corticosteroid 
administration 
531 
Nervous system 
disorders 
Cognitive 
disturbance 
Severe cognitive disability; 
significant impairment of 
work/school/life 
performance 
Both malignant glioma and its accepted 
therapies (for example radiation therapy) 
can produce impairment of work/school/life 
performance 
532 
Nervous system 
disorders 
Concentration 
impairment 
Severe impairment in 
attention or decreased level 
of concentration; limiting 
self care ADL 
Both malignant glioma and its accepted 
therapies (for example radiation therapy) 
can produce impairment in attention or 
decreased level of concentration that affects 
self care ADL 
536 
Nervous system 
disorders 
Dysesthesia 
Severe sensory alteration; 
limiting self care ADL 
Malignant glioma can produce alterations in 
sensory function limiting self care ADL 
545 
Nervous system 
disorders 
Headache 
Severe pain; limiting self 
care ADL 
Malignant glioma can result in intracranial 
pressure resulting in severe headaches 
567 
Nervous system 
disorders 
Pyramidal tract 
syndrome 
Severe symptoms; limiting 
self care ADL 
Malignant glioma can produce dysfunction 
of the pyramidal tract 
571 
Nervous system 
disorders 
Seizure 
Multiple seizures despite 
medical intervention 
Malignant glioma can produce multiple 
seizures despite medical intervention 
575 
Nervous system 
disorders 
Stroke 
Severe neurologic deficit 
Malignant glioma can produce stroke 
through its associated hypercoagulative 
state. Also, treatment for malignant glioma 
(for example bevacizumab, radiation) can 
result in stroke 
4.3 Supportive Care Guidelines 
Appropriate supportive care during the duration of the study includes the following: 
 
Bevacizumab or steroid administration for neurologic symptoms arising from increased edema or 
intracranial pressure, which are presumed mechanisms of action for M032, will be allowed, if deemed 
appropriate and safe in the determination of the investigator.  The increase in steroid dose will be kept to 
a minimum as patient neurologic condition permits. In general, UAB Neuro-oncology protocol is 
that Avastin is given every two weeks at a dose of 10/kg mg IV. Dosage adjustments or cessation of drug 

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may be made should adverse events (Uncontrolled hypertension, proteinuria, bleeding, thrombosis, etc.) 
occur and are made on a case-by-case basis by the prescribing neuro-oncologist. 
 
Proton pump inhibitors or H2 antagonists for control of steroid-induced gastric irritation 
 
Anti-epileptic medicines for control of partial or generalized seizures 
 
Post-operative neurological intensive care that is routine for the neurosurgical interventions involved in 
the administration of M032 
 
Other than a restriction on medications with anti-HSV activity (to be given only for the management of 
an adverse event), there are not any limitations on concomitant medications that patients may receive for 
other co-morbidities 
Management of adverse events is discussed in Section 5. 
4.4 Duration of Therapy/Study 
The therapeutic intervention in this trial involves the administration of a single infusion of M032.  For this 
reason it is more appropriate to define the duration of the study for each patient rather than the duration of therapy.  
All subjects participating in the study will be considered intent to treat and will be followed through all scheduled 
visits without concern for the total amount of study treatment received. 
In regard to the evaluations of treatment efficacy or the acceptability of other treatments for the malignant 
glioma, the study will continue as scheduled until one of the following criteria applies: 
 
Disease progression (as defined in Section 9) 
 
Patient withdrawal from the study 
Scheduled post-therapy safety evaluations as indicated in the study schedule will continue for every patient 
regardless of the response to M032.  These evaluations will stop only if one of the following criteria applies: 
 
Disease progression (as defined in Section 9) has rendered the patient not evaluable 
 
Patient withdrawal from the study 
4.5 Halting Rules for Study 
Should 2 or more Grade 3 or Grade 4 toxicities not thought to be related to tumor, prior therapy or expected 
to be related to HSV occur in the same dose level, enrollment will be suspended until an evaluation of these 
toxicities has been conducted by the trials Data Safety and Monitoring Board (DSMB). The DSMB will be the 
arbiter as to whether these toxicities can be attributed to M032 and if so, invoke a stopping rule, halting 
enrollment. If the DSMB should feel that these toxicities are not related to M032 administration, enrollment will 
be reinstituted as per its instructions. Should a particular DLT be evaluated by the DSMB and felt to be 1) 
consistent with M032 activity and 2) of acceptable duration and severity, the DSMB may decide to waive the 
DLT and/or enroll additional patients at this dose level due to the life-threatening nature of the underlying disease 
under study. 
 
The DSMB may interrupt study enrollment and entry at any time if medically indicated. 
 
If one (1) subject dies due to an event related to (caused by) the study intervention, study enrollment will 
be halted and enrollment may only begin after the DSMB reviews the data and the DSMB determines it 
is safe to continue.   
 
 
If two (2) subjects experience a Grade 4 toxicity using the NCTAE Toxicity Table, study enrollment will 
be halted and enrollment may only begin after the DSMB determines it is safe to continue.   
 
If two (2) instances of cytokine storms result in two instances of Grade 4 toxicity and/or uncontrolled 
corticoid steroids and acyclovir therapy are required, within the same cohort, study enrollment will be 
halted and enrollment may only begin after the DSMB reviews the data and the DSMB determines it is 
safe to continue.   

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 
If two (2) or more grade 3 toxicities not thought to be related to tumor, prior therapy or expected to be 
related to HSV occurring in the same dose cohort, enrollment will be suspended until an evaluation of 
these toxicities has been conducted by the trial’s DSMB. Enrollment may only begin after the DSMB 
reviews the data and the DSMB determines it is safe to continue. 
 
If two (2) or more Grade 4 neurologic toxicities not thought to be related to tumor, prior therapy or other 
treatment associated modalities occur in the same dose cohort, enrollment will be suspended until an 
evaluation of these toxicities has been conducted by the trial’s DSMB. Enrollment may only begin after 
the DSMB reviews the data and the DSMB determines it is safe to continue. 
 
5. EXPECTED ADVERSE EVENTS/DOSE MODIFICATIONS 
5.1 Expected Adverse Events Associated with M032 
Although M032 has not yet been used in humans, the adverse events it might produce are expected to be 
similar to the adverse events observed in the Phase I trials of a similar genetically-engineered herpes simplex 
virus, G207.  G207 caused no DLT and the adverse events that were observed following treatment were not 
thought to be attributable to the study drug.   
Adverse events occurring after the administration of G207 included the following, which are therefore 
expected to possibly occur following M032: 
 
 
asthenia 
 
hemiplegia 
 
dysphasia 
 
amnesia 
 
confusion 
 
depression 
 
nausea 
 
headache 
 
deep vein thrombosis 
 
somnolence 
 
decreased consciousness 
 
seizure 
 
leukopenia 
 
anemia 
 
abnormal erythrocytes 
 
peripheral edema 
 
cachexia 
 
urinary tract infection 
 
fever 
 
varicella zoster infection 
 
tumor progression 
 
pneumonia 
 
death 
 
abnormal mentation 
 
stroke 
 
increased liver 
transaminases 
 
pseudoprogression 
Other adverse events that might be expected based on the origin of M032 and on the patients’ disease include 
the following: 
 
 
hematoma 
 
encephalitis 
 
hepatitis 
 
disseminated HSV infection 
 
allergic response to M032 
 
nuchal rigidity 
 
photophobia 
 
autoimmune response to CNS tissues 
Any of the above adverse events will be considered a DLT, if possibly, probably or definitely attributable to 
M032 and if the severity of the adverse event is Grade 3 (in some situations) or Grade 4, as defined in Section 
4.2. 
Subjects will be observed closely for evidence of any adverse events.  After the delivery of M032, subjects 
will be observed in a Clinical Research Unit bed, a step down unit or the Neurosurgical ICU per the discretion of 
the surgeon.  The subjects’ vital signs (temperature, blood pressure, pulse, respiratory rate) and neurologic 
function and limited neurological exam will be monitored every hour for the first twelve hours, then every two 
hours for the next 12 hours, then every 4 hours for the next 24 hours. Thereafter, the frequency of monitoring 
will be determined by the attending physician(s) based on the medical condition of the patient. 
After discharge the subject will continue to be closely followed for evidence of adverse events, with 
outpatient follow-up evaluations scheduled at Day 10 and Months 1, 2, 3, 4, 5, 6, 9 and 12, or more often if 

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medically indicated. HSV detection will be performed at additional time points. The visits at months 2, 4 and 5 
will consist of sample collections and a study coordinator visit only.  Due to the complicated procedure for 
obtaining samples remotely at visits 2, 4 and 5, if a patient is unable to travel then, and if determined feasible by 
the PI, an attempt can be made to have the specimens collected by the patients’ referring physician and sent to 
UAB for analysis. 
The safety evaluation schedule is outlined in detail below in Section 8. Per FDA guidelines, subjects will be 
followed for survival. This will be achieved with monthly phone calls to subject. Additionally, when possible, 
subjects will be followed for emergence of any chronic or acute conditions that might be attributed to the study 
agent for up to 15 years during clinical visits with their standard of care practitioner. 
Most adverse events will be managed according to standard conventions.  General or specific neurologic  
worsening observed in the first several days after M032 administration could be due to edema, hydrocephalus, 
hematoma, or encephalitis.  Such problems occurring later may also be attributable to tumor progression.  When 
appropriate, an MRI will be done to help determine the cause of the neurologic changes. 
 
Cerebral edema/hydrocephalus: This commonly occurs in tumor patients post-operatively, and usually 
responds to standard measures for the treatment of increased intracranial pressure. 
 
Hematoma: PT, PTT, and platelet count will be obtained. A small hematoma may simply be watched and 
the patient treated as above for edema and then rescanned to exclude an enlarging lesion. A large 
hematoma or one associated with progressive neurologic deterioration may require operative evacuation. 
 
Encephalitis: Post-operative fever is not uncommon. However, fever >102F (with or without seizures) 
extending in duration >48 hours, in the presence of a waning Glascow Coma Scale and an increase in the 
area of hemorrhagic necrosis extending beyond the borders of the tumor on MRI are suggestive of viral 
encephalitis. If, in the opinion of the PI, it is safe and indicated, a cerebrospinal fluid sample will be 
obtained and analyzed by polymerase chain reaction (PCR) for evidence of HSV-1.  Otherwise, or if CSF 
results are not diagnostic, a stereotactic biopsy will be considered. This biopsy will be taken to assess for 
presence of M032 or wild-type HSV-1, as well as for histopathologic evidence of encephalitis. This will 
consist of a minimum of 2-3 needle core biopsies that will undergo standard hematoxylin and eosin 
(H&E) staining as well as immunostaining for HSV-1, leukocyte common  
antigen, glial fibrillary acidic protein (GFAP), and IL-12 immunostaining if possible.  High-dose antiviral 
therapy with intravenous acyclovir may be implemented in consultation with the Medical Monitor, Dr. 
John Gnann, and will be administered according to established method [37].  
 
Disseminated HSV Infection:  Disseminated multi-organ disease attributed to HSV is exceedingly 
uncommon, even in the immunocompromised host.  Such disease has been encountered in the newborn 
with disease identified as ‘disseminated multiorgan involvement’ and occurs in about 25% of newborns 
with this disease (incidence 1:10,000 deliveries).[38]  In adults, disseminated disease has been 
encountered in stem cell transplant recipients and, very rarely, pregnant women.[39]  In all 
circumstances, disseminated disease is associated with a systemic host response of fever, and findings 
compatible with sepsis, including hepatic dysfunction.  To monitor for these rare events, most 
importantly, serial physical examinations will be performed to assess for fever and symptoms of sepsis 
or hepatic dysfunction.  In addition, serial blood specimens will be obtained at Days 2, 10, 28 and monthly 
thereafter and cultured for infectious HSV.  Any positive cultures will be analyzed by PCR to 
discriminate between wild type HSV-1 and mutant M032 HSV.  Should a patient develop fever, rash, or 
unexplained elevations in liver function tests in conjunction with a concomitant increase of infectious 
HSV by plaque assay in blood specimens (exceeding prior values by 2 logs), patients will undergo a 21 
day course of treatment with acyclovir for presumed disseminated HSV infection. Parenthetically, it 
should be noted that oncolytic HSV therapy for metastatic colon carcinoma involving the liver led to 
detectable HSV DNA in the blood but in the ABSENCE of clinical symptomatology[40]; thus, increasing 
levels as well as fever will both be required to therapeutically  
intervene. Should 2 Grade 3 or 4 toxicities attributable to disseminated HSV infection occur in the same 
dose level, the prior dose level shall be declared the MTD and enrollment will be halted. 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 24 of 66 
Version 2.9 dated 20 Sep 2021 
 
Cytokine storm: although unlikely, this would not be entirely unexpected. Management of cytokine storm 
will focus on the pathology and symptoms.  Anti-inflammatory agents (for example corticosteroids) can 
minimize the cytokine response decreasing morbidity and mortality.  Should a cytokine storm or related 
toxicity occur, we will utilize antiviral therapy active against HSV (eg acyclovir) in combination with 
immunomodulatory agents to interrupt HSV replication and IL-12 production and effects, thereby 
reducing and/or eliminating morbidity and mortality.  Close observation of the subject for 24 hours after 
the crisis will be implemented (vital signs q hour, respiratory and cardiovascular support as needed, fluid 
maintenance as needed). 
5.2 Dosing Delays/Dose Modifications 
Because M032 is delivered as a single dose, there will not be any intra-patient dosing delays or dose 
modifications.   
5.3 Adverse Assessment and Management Assessment by PI/Clinician 
Neurologic complications are common in patients with malignant glioma and may be a result of tumor  
infiltration, mass effect, or other tumor associated complications; radiation administration and its associated direct 
and indirect toxicities; chemotherapy administration; and other medication and condition-related toxicities. Should 
2 or more neurologic toxicities that escalate to the level of a DLT and are not thought to be related to tumor, prior 
therapy or other treatment associated modalities occur in the same dose level, enrollment will be suspended until 
an evaluation of these toxicities has been conducted by the trials Data Safety and Monitoring Board (DSMB). The 
DSMB will be the arbiter as to whether these toxicities can be attributed to M032 and if so, invoke a stopping rule, 
halting enrollment. If the DSMB should feel that these toxicities are not related to M032 administration, enrollment 
will be reinstituted as per its instructions. 
Subjects will be monitored both at UAB and by their referring physician for any treatment-related toxicities 
until these have resolved back to their pretreatment baseline or < Grade 2.   
All unresolved toxicities will be followed regularly by the treating physician and/or research coordinator; the 
nurse coordinator will conduct follow-up weekly with the subject, family and/or referring physician (by phone if 
travel is too burdensome for the subject) until resolution of treatment-related toxicity to less than 
Grade 2 (or Grade 3 if that was the subject’s baseline). Additionally, subjects will be monitored monthly for the 
first 3 months following study drug administration, and will have the nurse-coordinator contact the subject, family 
and/or referring physician monthly for months 4-6. 
 
6. AGENT FORMULATION AND PROCUREMENT 
All agent materials and excipient solutions will be shipped to:  
Rachel Burell, Pharm.D. 
Research Pharmacist 
IDS Pharmacy 
North Pavilion, Room 3470 
1802 6th Avenue South 
Birmingham, AL 35249 
Phone: 205-934-7191  
Fax: 205-975-6647 
6.1 M032 Formulation and Storage 
HSV-M032 (NSC 733972), lot # L0909005, is supplied in sterile, labeled 1.0 mL single use glass vials 
containing 0.5 mL of HSV-M032 suspended in the storage buffer, Dulbecco’s Phosphate-buffered Saline (D-
PBS), 0.4M NaCl and 10% (w/v) glycerol, pH 7.4. The NExT Program recently reported the 48-month stability 
assay in which several reserved vials were retitered, resulting in a final calculated titer of 3.32 x 109 plaque-
forming units PFU/ml or 1.66 x 109 PFU per vial.  The vials should remain frozen at -70C or below until use.   
6.2 Availability 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 25 of 66 
Version 2.9 dated 20 Sep 2021 
M032 will be provided for clinical study under an Agreement between the NIH RAID program and the 
University of Alabama, Birmingham.  For specific shipment information/handling, refer to Appendix E.   
6.3 Excipient for Dilution of M032 
For dilution of M032, a specially formulated excipient solution will be used.  This excipient solution (Product 
Code 103382-50; Lot # 800612) has been purchased from Alanza Inc. (81 Glencameron Road, Thornhill, Ontario 
L3T 1N8, Canada).  The formulation is Phosphate Buffered Saline and 10% (w/v) of glycerol, pH 7.0-7.4 and is 
supplied sterile in 50 ml aliquots in amber serum bottles with snap-cap butyl rubber stoppers.   
6.4 M032 Dose Preparation 
All matters related to dose preparation and syringe set up for dose administration are performed in a sterile 
fashion. Prior to beginning infusion, the tubing that is to connect the syringes to the intracranial catheters is 
primed with sterile PBS + 10% glycerol (Alanza). All of the syringes containing the proper volume of  M032 
will be prepared at the same time for each three (3) hour period of delivery.  Syringes designated to be used 
beginning at three (3) hours should be kept at 4oC until dispensed to the patient.   
To prepare each dose, the vial(s) containing M032 should be thawed by removing it from the controlled 
access -60°C freezer and rubbing the vial gently between gloved hands until the last ice crystals have melted.  
The vial should then be placed on ice. Care should be taken to ensure that all the liquid is at the bottom of the 
vial before removing the cap. If it is suspected that the contents are on the side or top of the vial, the vial can be 
tapped gently on a flat surface. The cap must be removed carefully to avoid spilling or contamination.  The 
appropriate dose level of M032 will be removed from the vial and diluted in sterile diluent to the total volume.  
The final diluted M032 should be gently withdrawn into the syringe(s) for infusion. 
Once thawed, diluted and maintained at 4oC, the dose must be completely administered to the patient within 
sixteen hours of preparation.  The diluted virus has been shown to be stable at 4oC with no loss of activity for 9 
hours.   
Complete instructions for dose preparation are described in Appendices C and D.  
6.5 Precautions in Handling M032 
Sterile technique and Biosafety Level 2 precautions (gown, gloves, mask) will be rigorously followed while 
preparing the dose. The dose preparations will take place in a biosafety hood. 
6.6 Precautions in Disposal of M032 
All materials that have been in contact with M032 are considered infectious biohazards, and must be 
decontaminated or incinerated prior to disposal. Needles and syringes should be placed into a puncture-resistant, 
leak-proof container containing disinfectant. Instruments should be placed in disinfectant or swabbed with 
disinfectant soaked gauze sponges that are then discarded appropriately.  Instruments should be well cleaned and 
autoclaved.  All disposable materials that have been in contact with the vector must be incinerated or microwaved 
in an institutionally approved biohazard incinerator/microwave device before disposal. 
7. CORRELATIVE/SPECIAL STUDIES 
To address secondary objectives of the protocol, additional correlative studies will be performed on 
blood/serum, conjunctival secretions, saliva samples and tumor tissue samples.   
Blood Samples:  Blood samples (heparinized) will be taken at Baseline, on Days 0, 2, 3, 10, 28 and at Months 
3, 6, 9 and 12.  One ml of each sample will be submitted to the Clinical Virology Diagnostic Laboratory for HSV 
culture to determine the amount of virus shedding.  The balance of the blood sample will be separated by density 
gradient centrifugation to provide plasma (serum) and peripheral blood mononuclear cells (PBML).  Plasma will 
be tested to permit detection and quantification of HSV antibody titer pre-and post-inoculation via ELISA. 
Peripheral blood mononuclear leukocytes will be quantified for subsets and other leukocyte markers by FACS 
analysis, expression of activation signals (intracellular lymphocyte interferon-gamma levels) and lymphocyte 
transformation assays will also be performed to assess aspects of the TH1 response to treatment with M032. 
Plasma cytokine assays will be performed to quantify changes over time from baseline. Conjunctival, blood and 
saliva will be assessed for HSV shedding by standard culture methods.  Any positive cultures will be further 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 26 of 66 
Version 2.9 dated 20 Sep 2021 
characterized for wild-type or M032 HSV by PCR.   Any remnant blood will be stored for future brain tumor 
research. 
Tissue Samples: If available, remnant brain tissue from the biopsy will be snap-frozen in liquid nitrogen and 
stored for future research.  Should any patient with recurrent tumor after HSV treatment be determined to benefit 
from additional tumor resection for diagnosis or to ameliorate intractable symptoms caused by increased mass 
effect and intracranial edema, the tumor will be removed en bloc as much as possible and sectioned in 2-4mm 
thick slices perpendicular to the placement of the catheters.  Each section will be cut into quarters and each quarter 
will be labeled 1, 2, 3 or 4, processed and stored for future research as follows: 
Quarter 1:  Snap freeze in liquid nitrogen for immediate assessment of viral replication in vitro. 
Quarter 2:  Snap freeze in liquid nitrogen for RNA and DNA extraction and the following evaluations:  
• 
Quantitative PCR (vector replication in-vivo; compare number of copies to vector input)  
• 
RT-PCR to detect HSV polymerase mRNA (vector replication in-vivo)  
 
Confirmation and quantification of M032 presence by quantitative PCR of IL-12 recombinant DNA 
 
Confirmation and quantification of human IL-12 production by M032 by use of RT-PCR to the internal 
ribosomal entry site (IRES) sequence present between the p35 and p40 subunits of IL-12 in M032 
Quarter 3:  Snap freeze in liquid nitrogen for subsequent tissue homogenization and the following  
 
 
  
 
  evaluations: 
 
Confirmation and quantification of IL-12 production by use of a p70 ELISA kit after laser capture 
microdissection 
 
Quantification of in situ interferon- production by use of Multiplex Bead and ELISA kits 
 
Quarter 4:  Processed as routine histopathology in formalin for: 
 
Routine histopathology (H & E, GFAP, MIB, etc.) 
 
Immunohistochemistry for HSV-1, glycoprotein C (gC) when possible 
 
Characterization of the nature of the inflammatory infiltrate by reaction with monoclonal antibodies for 
CD4, CD8 and macrophage markers 
Subsequent 2-4mm thick Slices if available): 
 
Process 2 quadrants of each slice in formalin as routine histopathology; may be used for further 
characterization of the inflammatory infiltrate as needed 
 
Snap freeze 2 quadrants of each slice in liquid nitrogen; may be used for further studies, such as laser 
capture microdissection, RNA and DNA extraction as needed, etc. 
 
Schematic for Processing of Tumor Tissue 
Quarter 1
PCR and
RT-PCR for
HSV
Quarter 2 
PCR and 
RT-PCR for 
murine 
IL-12
Quarter 4
IHC for 
CD4, CD8,
and 
macrophages
Quarter 3 
ELISA for 
murine Il 
-12
A 
B
C
3-5 mm 
Catheter
Tissue 
D 
E
 
 
Quarter 1 
Laser capture/ 
ELISA IL-12, 
IFN- gamma 
Quarter 2 
q-PCR for vector, Il-12 
RT-PCR for IRES/IL-
12, polymerase 
Quarter 4 
Histopath/ 
 H & E, 
 IHC leuckocytes, gC 
Quarter 3 
Culture for 
infectious virus 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 27 of 66 
Version 2.9 dated 20 Sep 2021 
8. STUDY CALENDAR 
 
 
Pre-Study               
(within 2 weeks prior 
to the administration 
of M032) 
Day 0 
Day1 
Day 2 
Day 3 
Day 10 ( ± 3 days) 
Day 28 ( ± 4 days) 
Month 2 ( ± 12 days) 
Month 3 ( ± 12 days) 
Months 4, 5             
( ± 12 days) 
Months 6, 9, 12           
( ± 12 days) 
Unscheduled20 
Informed consent 
X1 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/ Exclusion 
Criteria 
X 
 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
Pregnancy test-serum4 
X 
 
 
 
 
 
 
 
 
 
 
 
Medical history 
X 
 
 
 
 
 
 
 
 
 
 
 
Concurrent meds 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Complete physical exam 
(including  wound 
assessment Days 1-10) 
X 
 
X 
X17 
X17 
X 
X 
 
X 
 
X 
X21 
Vital signs 
X 
X 
X2 
X2 
X2 
X 
X 
 
X 
 
X 
X21 
KPS 
X 
 
 
 
X 
X 
X 
 
X 
 
X 
X21 
CBC w/diff, plts 
X 
 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
Serum chemistry3, 
PT/INR, PTT 
X 
 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
HIV serology 
X 
 
 
 
 
 
 
 
 
 
 
 
EKG 
X 
 
 
 
 
 
 
 
 
 
 
 
CXR (AP and lateral) 
X 
 
 
 
 
 
 
 
 
 
 
 
Urinalysis with micro 
X 
 
 
 
 
 
 
 
 
 
 
 
Adverse event 
evaluation11, 12 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
MRI5 
X13 
X 
 
 
X 
 
X 
 
X 
 
X 
X21 
Neurologic exam 
X 
X 
X6 
X6 
X6 
X 
X 
 
X 
 
X 
X21 
HSV Ab titer7 
X 
 
 
 
 
X 
X 
 
X 
 
X 
X 
HSV detection (saliva, 
conjunctival secretions, 
blood)8 
X 
 
 
X18 
X18 
blood 
only 
X 
X 
X 
X 
X 
X 
X 
IL-12 detection (serum)9 
X 
 
 
X18 
X18 
X 
X 
 
X 
 
X 
X 
Blood sample for LTA, 
Elispot 
X 
 
 
X18 
X18 
X 
X 
 
X 
 
X 
X 
IFN gamma assay 
X 
 
 
X18 
X18 
X 
X 
 
X 
 
X 
X 
Blood to be stored for 
future immune studies 
X 
X15  
 
X18 
X18 
X 
X 
 
X 
 
X  
X  
Head CT 
 
 
 
X10 
 
 
 
 
 
 
 
 
 
 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 28 of 66 
Version 2.9 dated 20 Sep 2021 
 
Pre-Study               
(within 2 weeks prior 
to the administration 
of M032) 
Day 0 
Day1 
Day 2 
Day 3 
Day 10 ( ± 3 days) 
Day 28 ( ± 4 days) 
Month 2 ( ± 12 days) 
Month 3 ( ± 12 days) 
Months 4, 5             
( ± 12 days) 
Months 6, 9, 12           
( ± 12 days) 
Unscheduled20 
Biopsy, Catheter 
placement 
 
X 
 
 
 
 
 
 
 
 
 
 
Prophylactic AED 
X 
X 
X 
X 
X 
X 
X16 
 
 
 
 
 
M032 Administration 
 
 
X 
 
 
 
 
 
 
 
 
 
Rickham/Ommaya 
reservoir aspiration19 
X 
X 
X 
X 
X 
X 
X 
 
X 
 
X 
X21 
 
1. Will be obtained prior to study screening procedures. 
2. Frequency post initiation of infusion: vital signs will be every hour for 12 hours post initiation, every 2 hours for the 
next 12 hours, then every 4 hours for the next 24 hours. 
3. Includes sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, calcium, phosphorus, total protein, uric 
acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, total bilirubin, cholesterol, triglyceride. 
4. Only women of child-bearing potential. If necessary, can be repeated during the course of the study. 
5. All MRI scans will be performed as prescribed by the Principal Investigator. 
6. Neurological exam includes assessment of cranial nerve abnormalities, gait abnormalities (when appropriate), sensory 
abnormalities, muscle strength, and level of consciousness. This will be evaluated every hour for 12 hours post initiation, 
every 2 hours for the next 12 hours, then every 4 hours for the next 24 hours. 
7. By ELISA; neutralizing antibody assays may also be performed. 
8. Samples will be evaluated by PCR and culture; quantitative PCR may be performed on positive samples. 
9. IL-12 detection (serum) by ELISA.   
10. After catheter placement but before start of infusion; may also take place on Day 1. 
11. All AEs will be assessed throughout the course of the study, but specific CRFs will only be required at the time points 
delineated in the above schedule. 
12. All unresolved toxicities be followed regularly by the treating physician and/or research coordinator; the nurse 
coordinator will conduct follow-up weekly with the subject, family and/or referring physician (by phone if travel is too 
burdensome for the subject) until resolution of treatment-related toxicity to less than Grade 2 (or Grade 3 if that was the 
subject’s baseline). 
13. MRI’s that have been obtained within 30 days of pre-study visit may be used at the discretion of the PI. 
14. Sample will be stored for future brain tumor research. 
15. To be obtained post-op. 
16. Prophylactic AED’s may be discontinued at the Day 28 visit. 
17. Wound examination only. 
18. Acceptable up to 2 days after 
19. Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the more central 
portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 2-10cc)  from the catheter 
for laboratory studies. These specimens will be obtained at either the pre op clinic visit or the day of surgery, as well 
as up to daily during the inpatient hospitalization (per the surgeon’s discretion), and again at post op visit day 10, Day 
28, and Months 3,6,9 and 12. 
20. If a participant is to miss a regularly scheduled research follow-up visit, an unscheduled visit can be conducted to 
capture research data at an otherwise standard of care clinic visit.  
21. Collected only if done during an otherwise non-research visit (i.e., unscheduled visit). 
 
8.1 Study Procedures 
 
 Screening Phase: (within 2 weeks prior to Treatment Phase) 
 
Obtain informed consent before any study-related procedures are performed 
 
Review Inclusion/Exclusion Criteria for eligibility 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 29 of 66 
Version 2.9 dated 20 Sep 2021 
 
Collect the following information: 
 
Demographics 
 
Medical history  
 
Medications currently being taken 
 
Obtain vital signs (pulse, blood pressure, respiratory rate, temperature, height and weight) 
 
Perform a complete physical examination  
 
Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate),  sensory abnormalities, muscle strength, and 
level of consciousness  
 
Complete the Karnofsky Performance Scale (KPS) 
 
 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, 
calcium, phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, 
total bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HIV Serology 
 
Pregnancy test (if applicable) 
 
HSV antibody titer 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay  
 
Immune studies - to be stored for future research 
 
Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
 
Collect urine sample for urinalysis and microscopic examination 
 
Obtain: 
 
ECG 
 
Chest x-ray 
 
MRI - MRI’s that have been obtained within 30 days of pre-study visit may be used at the 
discretion of the PI. 
 
All patients will be required to be placed on an anticonvulsant at the time of screening; those patients 
already on an anti-convulsant (anti-epileptic drug/AED) will have a second anti-convulsant added per 
the discretion of the investigator. This anti-convulsant will be continued for a minimum of 28 days 
unless the patient develops a reaction to the medication. If this develops, all attempts will be made to 
implement an alternative agent. 
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
 
Day 0 (biopsy and catheter placement)  
 
Admit to the hospital for biopsy surgery and if eligible, catheter placement 
 
Review medications  
 
AE monitoring 
 
Obtain vital signs (pulse, blood pressure, respiratory rate, temperature and weight) prior to surgery 
 
Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate), sensory abnormalities, muscle strength, and level 
of consciousness  
 
Obtain a stereotactic MRI to determine site for the biopsy and catheter placement 
 
Perform the biopsy and, if eligibility criteria are met, place and label up to 4 catheters 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 30 of 66 
Version 2.9 dated 20 Sep 2021 
 
Prime each catheter with 0.4 ml sterile saline solution and cap the catheters 
 
Admit to the Neurosurgery Intensive Care Unit for routine post-biopsy care and observation 
 
Obtain CT scan to ensure the catheters are in the proper position. (May be completed on Day1 prior to 
drug administration 
 
Collect blood for the following:   
 
Immune studies - to be stored for future research – collect post-op 
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
 
Day 1 (drug administration): 
 
If subject’s condition is stable, transfer to a Clinical Research Unit inpatient bed for drug infusion and 
monitoring 
 
Review medications  
 
Perform a complete physical exam including wound assessment 
 
If not completed on Day 0, obtain a CT scan to ensure the catheters did not move and are still in the 
proper place 
 
Obtain pathology report and verify diagnosis 
 
 
Administration of study drug which will take approximately 6 hours (may last up to 12 hours)  
 
Study Drug Administration 
 
Acquisition of study drug and diluent 
 
Store syringes in refrigerator (4° C) until needed 
 
Flush infusion tubing with diluent provided by pharmacy 
 
Connect syringe with study drug to infusion tubing 
 
Prime with 4.6 ml of the diluted virus preparation making sure that any excess study drug is 
disposed according to Biosafety Level 2 procedure  
 
Connect tubing to catheter 
 
Place each syringe in syringe pump  
 
Repeat for each active catheter 
 
Program pump(s) to flush saline solution at a rate of 0.4 ml per catheter for 35 minutes  
 
Re-program pump(s) to deliver study drug as follows: 
 
Four (4) active catheters – 0.1ml/hour 
 
Three (3) active catheters – 0.13 ml/hour 
 
Two (2) active catheters – 0.2 ml/hour 
 
One (1) active catheter – 0.4ml/hour 
 
Begin infusion 
 
After the 3 hour study drug infusion is complete (study drug syringe will not be empty) and 
prior to disconnecting first infusion tubing from catheter, flush  second infusion tubing with 
diluent supplied by pharmacy 
 
Obtain second syringes from refrigerator 
 
Connect syringe to infusion tubing and prime with 4.6 ml of the diluted virus preparation 
 
Disconnect first tubing from catheter and connect second tubing 
 
Place syringe in syringe pump  
 
Repeat for each active catheter 
 
Program pump(s) to deliver study drug at 
 
Four (4) active catheters – 0.1ml/hour 
 
Three (3) active catheters – 0.13 ml/hour 
 
Two (2) active catheters – 0.2 ml/hour 
 
One (1) active catheter – 0.4ml/hour  

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
Page 31 of 66 
Version 2.9 dated 20 Sep 2021 
 
Begin infusion If there is any problems with the catheters during the infusion, the PI 
and/or Co-PI are to be notified as well as the Study Coordinator 
 
Obtain vital signs (pulse, blood pressure, respiratory rate and temperature) according to the following 
schedule: 
 
Before initiation of study drug   
 
Every hour for 12 hours post initiation 
 
Every 2 hours for the next 12 hours 
 
Every 4 hours for the next 24 hours 
 
Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate),  sensory abnormalities, muscle strength, and level 
of consciousness: 
 
Before initiation of study drug 
 
Every hour for 12 hours post initiation 
 
Every 2 hours for the next 12 hours 
 
Every 4 hours for the next 24 hours 
 
AE monitoring  
 
After infusion is completed (study drug syringe will not be empty), the catheters will be disconnected 
from the infusion tubing and capped off.  The catheters will remain in place for 6-18 hours and then 
removed. 
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
Day 2: 
 
Continuation of vital signs and neurological examinations (see Day 1 for schedule) 
 
Wound examination 
 
Review of medications  
 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, 
calcium, phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, 
total bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay 
 
Immune studies - to be stored for future research 
 
Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
 
AE monitoring  
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
 
Day 3: 
 
Continuation of vital signs and neurological examinations (see Day 1 for schedule) 
 
Wound examination 
 
Review of medications 
 
Complete the Karnofsky Performance Scale (KPS) 
 
Obtain MRI scan  prior to discharge 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
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 
Collect blood for the following: 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay 
 
Immune studies - to be stored for future research 
 
AE monitoring 
 
Discharge if subject is stable enough to leave the hospital 
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
 
Day 10 ± 3 days: 
 
Review medications 
 
AE monitoring 
 
Obtain vital signs (pulse, blood pressure, respiratory rate, temperature and weight)   
 
Perform a complete physical exam 
 
Wound examination 
 
Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate),  sensory abnormalities, muscle strength, and level 
of consciousness  
 
Complete the Karnofsky Performance Scale (KPS) 
 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, 
calcium, phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, 
total bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HSV antibody titer 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay 
 
HSV detection 
 
Immune studies - to be stored for future research 
 
Collect samples of  conjunctival (eye) secretions and saliva for HSV detection 
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
 
Day 28 ± 4 days: 
 
Review medications 
 
AE monitoring 
 
Obtain vital signs (pulse, blood pressure, respiratory rate, temperature and weight)   
 
Perform a complete physical exam 
 
Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate),  sensory abnormalities, muscle strength, and level 
of consciousness  
 
Complete the Karnofsky Performance Scale (KPS) 

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 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, 
calcium, phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, 
total bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HSV antibody titer 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay 
 
Immune studies - to be stored for future research 
 
Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
 
Obtain MRI scan   
 
Discontinue prophylactic AED medication (if applicable) 
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
 
Month 2 (study coordinator visit): 
 
Review medications 
 
AE monitoring 
 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, 
calcium, phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, 
total bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HSV detection 
 
Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
 
Month 3: 
 
Review medications 
 
AE monitoring 
 
Obtain vital signs (pulse, blood pressure, respiratory rate, temperature and weight)   
 
Perform a complete physical exam 
 
Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate),  sensory abnormalities, muscle strength, and level 
of consciousness  
 
Complete the Karnofsky Performance Scale (KPS) 
 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, 
calcium, phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, 
total bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HSV antibody titer 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay 
 
Immune studies - to be stored for future research 

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 
Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
 
Obtain MRI scan   
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
 
Months 4 and 5 (study coordinator visit): 
 
Review medications 
 
AE monitoring 
 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, calcium, 
phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, total 
bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HSV detection 
 
Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
 
Months 6 and 9: 
 
Review medications 
 
AE monitoring 
 
Obtain vital signs (pulse, blood pressure, respiratory rate, temperature and weight)   
 
Perform a complete physical exam 
 
Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate),  sensory abnormalities, muscle strength, and level 
of consciousness  
 
Complete the Karnofsky Performance Scale (KPS) 
 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, calcium, 
phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, total 
bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HSV antibody titer 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay 
 
Immune studies - to be stored for future research 
 
Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
 
Obtain MRI scan 
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12   
 
Month 12 (end of study): 
 
Review medications 
 
AE monitoring 
 
Obtain vital signs (pulse, blood pressure, respiratory rate, temperature and weight)   

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 
Perform a complete physical exam 
 
Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate),  sensory abnormalities, muscle strength, and level 
of consciousness  
 
Complete the Karnofsky Performance Scale (KPS) 
 
Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, calcium, 
phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, total 
bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 
 
HSV antibody titer 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay 
 
Immune studies - to be stored for future research 
 
Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
 
Obtain MRI scan   
 
Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 
2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at either the pre op 
clinic visit or the day of surgery, as well as up to daily during the inpatient hospitalization (per the 
surgeon’s discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12 
 
Unscheduled Visit: 
 
The following data will be collected at a standard of care follow-up visit only if it has been collected 
for standard of care purposes (that is: these procedures will not be conducted for research purposes 
only): 
o Collect vital signs (pulse, blood pressure, respiratory rate, temperature and weight), if done as SOC 
o Collect a complete physical exam data, if done as SOC 
o Perform a neurological (nervous system) examination to include assessment of cranial nerve 
abnormalities, gait abnormalities (when appropriate),  sensory abnormalities, muscle strength, and 
level of consciousnesss, if done as SOC 
o Collect the Karnofsky Performance Scale (KPS), if done as SOC 
o Obtain MRI scan, if done as SOC 
 
 
The following procedures will be performed for research purposes only: 
o Review medications 
o AE monitoring 
o Collect samples of conjunctival (eye) secretions and saliva for HSV detection 
o Should a patient have an access device (eg., Rickham or Ommaya reservoir) that accesses either the 
more central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible 
(expected, 2-10cc)  from the catheter for laboratory studies. These specimens will be obtained at 
either the pre op clinic visit or the day of surgery, as well as up to daily during the inpatient 
hospitalization (per the surgeon’s discretion), and again at post op visit day 10, Day 28, and 
Months 3,6,9 and 12 
o Collect blood for the following:   
 
CBC with diff and platelets 
 
Serum Chemistry (sodium, potassium, chloride, bicarbonate, glucose, BUN, creatinine, calcium, 
phosphorus, total protein, uric acid, GGT, AST/ALT, CK, LDH, alkaline phosphatase, total 
bilirubin, cholesterol, triglyceride) 
 
PT/INR, PTT 

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 
HSV antibody titer 
 
HSV detection 
 
IL-12 detection 
 
LTA Elispot 
 
IFN gamma assay 
 
Immune studies - to be stored for future research 
 
 
9. MEASUREMENT OF EFFECT 
Although response is not the primary endpoint of this trial, patients with measurable disease will be assessed 
by standard criteria. 
9.1 Definitions 
Although the Response Evaluation Criteria in Solid Tumors (RECIST) Committee has proposed new 
international criteria for the evaluation of response and progression of solid tumor, unidimensional measurements 
alone have not yet been sufficiently validated for the evaluation of malignant gliomas. The MacDonald criteria 
will be used for evaluation of treatment responses which is more appropriate for patients who have failed surgery, 
radiation and chemotherapy prior therapy than recently described RANO criteria which is more appropriate for 
newly diagnosed patients with high-grade, malignant brain tumors [39]. 
9.1.1 
Measurable Disease 
Measurable lesions are defined as those that can be accurately measured in at least one dimension 
(longest diameter to be recorded) as >10 mm with MRI scan.  All intracranial malignant glioma is therefore 
measurable disease.  All tumor measurements must be recorded in millimeters (or decimal fractions of 
centimeters). 
9.1.2 
Non-measurable Disease 
This type of lesion is not applicable in this study. 
 
9.1.3 
Target Lesions 
All intracranial lesions that are likely manifestations of the patient’s malignancy will be considered 
target lesions. 
9.1.4 
Non-target Lesions 
This type of lesion is not applicable in this study. 
9.2 Guidelines for Evaluation of Measurable Disease 
All measurements should be taken and recorded in metric notation using a ruler or calipers, or digitized 
method.  All screening evaluations should be performed as closely as possible to the beginning of treatment and 
never more than 4 weeks before the beginning of the treatment. 
Because all of the patients enrolled in this study will have received prior radiation therapy, tumor lesions that 
are situated in a previously irradiated area will be considered measurable. 
The same method of assessment and the same technique should be used to characterize each identified and 
reported lesion at baseline and during follow-up. Imaging-based evaluation is the only means of assessing the 
antitumor effect of this treatment for this disease.   
9.3 Response Criteria 
The following criteria are based upon the volume changes that correspond to the definitions proposed by the 
MacDonald criteria.  For the purposes of this study, the objective response of each patient’s disease to  
M032 will be evaluated by the comparison of a baseline MRI scan to follow-up MRI.  For those patients receiving 
intratumoral inoculation, the screening MRI will serve as the baseline MRI, and follow-up scans will  
be done at study Day 3, Day 28, Month 3, Month 6, Month 9 and Month 12.  To prevent the introduction of 
observer bias, a software package which determines tumor areas and volumetrics via assessment of pixel intensity 
will be utilized to compare pre-and post-treatment images. 

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9.3.1 Evaluation of Target Lesions 
“Response” is defined as follows: 
 
Complete Response (CR) - Disappearance of all treated enhancing tumor on MRI scan, off 
steroids, and neurologically stable or improved. 
Partial Response (PR) - greater than 50% reduction in the treated enhancing tumor on MRI 
scan, stable or reduced steroid dose, and neurologically stable or improved. 
Progressive Disease (PD) - greater than 25 % increase in the treated enhancing tumor on 
MRI scan, stable or increased steroid dose, and neurologically stable or worse. 
Stable Disease (SD) - all other situations 
Some patients on this trial may be taking dexamethasone and/or bevacizumab. Those who have decreased 
the dose of one of these medications to one lower than that utilized at the time of the previous MRI AND this 
dose decrease has occurred within two weeks of the MRI shall not be determined to have progressed even if 
the MRI meets the criteria for PD above, but rather that MRI shall be considered non-evaluable for response. 
Similarly, should an increase in steroid dosage or bevacizumab dosage occur within two weeks of an MRI 
compared to the prior MRI, the patient will not be eligible for consideration of a CR or PR but instead that 
MRI will be considered non-evaluable. 
 
Note that since this is considered an immunotherapy trial, the appearance of lesion(s) does not necessarily 
indicate PD. 
9.3.2 
Evaluation of Non-target Lesions 
Because all intracranial lesions will be considered target lesions, this assessment is not applicable to 
this study. 
 
9.3.3 
Evaluation of Best Overall Response 
The best overall response is the best response recorded from the start of the treatment until disease 
progression/recurrence (taking as reference for progressive disease the smallest measurements recorded since 
the treatment started).  The patient's best response assignment will depend on the achievement of both 
measurement and confirmation criteria (see section 9.3.1). 
Note: In some circumstances, it may be difficult to distinguish residual disease from normal tissue.  
When the evaluation of complete response depends on this determination, it is often recommended that the 
residual lesion be investigated (biopsy) before confirming the complete response status.  Because this disease  
would require a brain biopsy in this situation, with potential increased risk to the patient, patients with an 
ambiguous complete response status will only undergo histological confirmation if deemed necessary for 
additional treatment interventions.  If tumor is not the predominant feature of biopsy specimen, e.g., if only 
necrosis is present, or inflammation or gliosis are prominent, then progression will not be deemed to have 
occurred. [40] 
 
 
9.3.4 
Pseudoprogression 
Pseudoprogression is a well-known entity in oncolytic viral therapy of glioma. Currently, the 
immunotherapy Response Assessment in Neuro-Oncology (iRANO) group is proposing criteria for 
determination of pseudoprogression in immunotherapy trials which are not yet published. [40] We will utilize 
the following criteria to address this phenomenon in this trial : If tumor appears to enlarge or demonstrates 
increased enhancement by MRI consistent with PD, patients will undergo repeat imaging and clinical 
assessments at 3 months to determine whether the changes demonstrate true progression or 
pseudoprogression. Note that the investigator may obtain an MRI prior to this 3 month interval as indicated. 
Should significant neurologic deterioration occur that cannot be ascribed to tumor or pseudoprogression 
related events such as seizures or medication changes (e.g., steroid tapers) the patient will be considered to 
have progressed. During the interval, the patient may be watched or treated with bevacizumab and/or 
dexamethasone at the discretion of the investigators. If the patient improves with bevacizumab (to be used 

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preferentially at a suggested dose of 10mg/kg IV, every two weeks, for three months; previous tumor 
progression on bevacizumab does not preclude its use herein) and/or additional steroid administration (to be 
used if bevacizumab is contraindicated or insufficient) and the MRI changes in lesion size/and or 
enhancement also improves, the patient will be determined to have not progressed but to have suffered 
pseudoprogression, and will continue follow-up within the trial under the previously defined schedule. 
Should neurologic symptoms and/or imaging changes progress despite steroid administration on follow-up 
imaging, the patient will be determined to have progressed and the date of progression shall be assigned to 
the date of the initial scan that demonstrated findings consistent with PD as defined above.  
9.4 Confirmatory Measurement/Duration of Response 
9.4.1 
Confirmation 
To be assigned a status of PR or CR, changes in tumor measurements must be confirmed by the next 
scheduled MRI after the criteria for response are first met.  In the case of SD, follow-up measurements must 
have met the SD criteria at least once after study entry at a minimum interval of 12 weeks. 
9.4.2 
Duration of Overall Response 
The duration of overall response is measured from the time measurement criteria are met for CR or 
PR (whichever is first recorded) until the first date that recurrent or progressive disease is objectively 
documented (taking as reference for progressive disease the smallest measurements recorded since the 
treatment started).  The duration of overall CR is measured from the time measurement criteria are first met 
for CR until the first date that recurrent disease is objectively documented. 
 
9.4.3 
Duration of Stable Disease 
Stable disease is measured from the start of the treatment until the criteria for progression are met, 
taking as reference the smallest measurements recorded since the treatment started. 
9.5  Progression-Free Survival 
 
Because this study is a non-randomized phase 1 trial that will enroll relatively few patients, 
progression-free survival will be reported as a secondary endpoint. 
10. REGULATORY AND REPORTING REQUIREMENTS 
Adverse events (AE) will use the descriptions and grading scales found in the revised NCI Common Toxicity 
Criteria for Adverse Events (CTCAE).  This study will utilize the CTCAE version 4.0 for adverse event reporting.  
All appropriate treatment areas will have access to a copy of the CTCAE version 4.0.  A table showing the 
expected adverse events associated with M032 and the related IMT terms can be found in Appendix A.   
10.1 
Adverse Event Reporting (AE; formerly known as Adverse Drug Reaction) 
10.1.1 Adverse Event 
 
An adverse event (AE) is defined as any untoward medical occurrence associated with the use of a drug 
in humans, whether or not considered drug related. An AE can be any unfavorable and unintended sign 
(e.g., abnormal laboratory finding), symptom, or disease temporally associated with the use of a drug, 
without any judgment about causality. An AE can arise from any use of the drug and from any route of 
administration, formulation, or dose, including an overdose. 
  An illness or condition present at study entry is considered a pre-existing condition and will not be 
considered an AE unless the pre-existing condition worsens during the course of the study.  This change 
may be considered an AE. An AE that occurs after signing informed consent but prior to the administration 
of study drug will be considered not related to study treatment. 
  AE’s that have a grade increase to 4 or 5 will be considered an SAE and reported as required. 
 
 
All AE’s, irrespective of relationship to study drug, which occur or worsen after signing of consent 
form until 30 days after administration of study drug, must be reported and documented on the AE Case 

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Report Form (CRF).  All medications given for an AE will be reported on the Concomitant Medication 
CRF. 
 
 
Hospitalizations that are expected due to the disease and those that are unrelated to the study drug or 
disease will be reported as AE’s unless it meets the requirements for a serious adverse event (SAE) which 
can be found in Section 10.1.2.  Hospitalizations due to merely diagnostic reasons, examinations as a 
matter of routine or planned surgery and due to social indication do not represent a hospitalization in the 
sense of the term “serious”. 
 
An AE is considered unexpected if it is not listed in Section 5.1. 
10.1.2 Serious Adverse Event Expedited Reporting Guidelines – Phase 1 studies with investigational 
agents: 
 
A serious adverse event (SAE) is defined as: 
 
 
Unexpected AE  
 
For grade 1, no expedited reporting required 
 
For grade 2, the investigator will be unbiased in assessing the relationship of the 
event and test article and discuss the possible relatedness to treatment with the 
Medical Monitor. If found to be possibly, probably or definitely related to study 
drug, an expedited report will be submitted within 7 working days. 
 
For grade 3, the investigator will be unbiased in assessing the relationship of the 
event and test article and discuss the possible relatedness to treatment with the 
Medical Monitor. If found to be possibly, probably or definitely related to study 
drug, an expedited report will be submitted by phone within 24 hours with 
expedited report to follow within 7 working days. 
 
For grade 4-5, regardless of relationship to study drug, a report will be submitted 
by phone within 24 hours with expedited report to follow within 7 working days. 
 
AE whose grade has increased to grade 3 or 4 
 
Adverse Event and IND safety reports will be filed in a timely manner with institutional 
authorities (IRB, IBC) as well as with the FDA and NIH/ORDA.  
 
All serious adverse events (Grade 3 or 4 toxicities) will be reported by fax, e-mail or phone 
within 24 hours and a written expedited report filed within seven days. Additionally, unexpected 
Grade 2 or Grade 3 toxicities will require a written expedited report within seven 
 
days and Grade 3 unexpected adverse events will also require a fax, email, or phone call to the 
UAB IRB, the Neuro-Oncology CTMC, CCC PRC, FDA, and OBA within 24 hours.   
 
For events that are related, unexpected and serious, reporting will be reported to the FDA within 
7 days of knowledge of the event. 
 
A list of agent-specific expected adverse events can be found in Appendix A. 
 
Any adverse event requiring an expedited report will also be reported immediately to the Medical 
Monitor and the investigator’s Institutional Review Board (IRB). 
UNEXPECTED EVENT 
EXPECTED EVENT 
GRADES 2 – 3 
Attribution of Possible, Probable or Definite 
GRADES 4 and 5 
Regardless of Attribution 
GRADES 1 - 3 
GRADES 4 and 5 
Regardless of Attribution 
 
Grade 2 - Expedited report within 7 working days. 
 
Grade 3 - Report by phone to IDB within 24 hrs. 
Expedited report to follow within 7 working days. 
 
Report by phone to IDB 
within 24 hrs. Expedited 
 
Adverse Event 
Expedited 
 
Report by phone to IDB within 
24 hrs.  Expedited report to 
follow within 7 working days. 
 

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10.1.3 AE and SAE Summary Table 
10.1.4  Forms 
We will utilize MedWatch for adverse event reporting.  A copy of this form is provided in Appendix 
F. 
10.1.5  Secondary Malignancies 
Investigators are required to report secondary malignancies occurring on or following treatment on 
NCI sponsored protocols using the form noted above.  Exception:  Cases of secondary AML/MDS are to 
be reported using the NCI/CTEP Secondary AML/MDS Report Form. 
10.2 
Data Reporting 
This study will be under the supervision of an external Data and Safety Monitoring Committee without 
conflicts of interest who will be appointed and chartered to oversee the review of all adverse events and efficacy 
determinations. The reviews will be completed quarterly and in real time.  The committee will also oversee the 
clinical trial data analysis and publication of study results. The DSMB will be comprised of a medical infectious 
disease expert, a neurosurgeon familiar with oncolytic virus therapy for brain tumors, a neuro-oncologist familiar 
with oncolytic virus therapy for patients with brain tumors and an expert in the use of current molecular biology 
techniques for the detection and quantification of Herpes Simplex Virus in clinical specimens.  The Medical 
Monitor is a medical infectious disease expert.  Any member who resigns or otherwise leaves service on the 
DSMB will be replaced by an expert with equivalent experience and expertise and without any potential conflicts.  
The Data and Safety Monitoring Board for this Phase I study will utilize the appended NCI-approved Data and 
Safety Monitoring Plan (Appendix G). 
10.3 
CTEP Multicenter Guidelines:   
Not applicable. 
10.4 
Cooperative Research and Development Agreement (CRADA)/Clinical Trials Agreement (CTA):   
Not applicable. 
11. STATISTICAL CONSIDERATIONS 
We will implement the CRM design (the dosing range is described previously in this protocol) in a manner 
similar to that of Geletneky et al. [43] We will define the primary criterion for dose modification as the relative 
frequency of a DLT at a given dose level. A dose will be considered as tolerable if the probability of toxicity is 
<33%. The Recommended Phase 2 Dose (RP2D) will be the dose level with the highest posterior probability of 
having a toxicity of ≤33%. The Optimal Biologic Dose (OBD) will be the RP2D that is obtained by enrolling the 
fewest number of evaluable patients possible. The posterior probability for toxicity will be determined using a 
logistic model, where the toxicities will be parameterized as probabilities in the model. When a patient enters the 
study, the dose level will be determined by calculating the posterior probability for each dose level to be tolerable. 
The highest dose level that is tolerable will be allocated. Regarding the dose escalation scheme, a conventional 
three-at-one [44] design will be used as long as no DLT is observed. After the first DLT is observed, one 
additional patient will be enrolled at the same dose before the dose will be determined as the RP2D using the 
logistic model. The highest dose with a median probability of at least 33% will be applied as the current RP2D 
to the subsequent patient, following the CRM [45]. If the RP2D is calculated as dose level -1 (i.e. 1 X 104 PFU), 
the trial will end with no safe dose established. 
11.1 
Study Design/Endpoints  
This study is an open-label, dose escalating phase 1 study of the safety of intracranial administration of 
 
(Grade 1 – Adverse Event Expedited Reporting NOT 
required.) 
report to follow within 7 
working days. 
 
This includes deaths within 
30 days of the last dose of 
treatment with an 
investigational agent. 
Reporting NOT 
required. 
This includes deaths within 30 
days of the last dose of 
treatment with an 
investigational agent. 

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M032, a genetically engineered HSV-1 expressing IL-12.  For this reason, the primary endpoints of this study is 
to determine the safety and tolerability of a single 12 hour stereotactic intracerebral infusion of escalating doses 
of M032 virus, and to determine the maximally tolerated dose (MTD) of M032.  The dose escalation scheme and 
the definition of the MTD are found in Section 4.2.  Because M032 is administered as a single dose (although 
that single dose may be delivered via different catheters), intra-patient dose escalations will not be feasible. 
Demographic Analysis: Demographic and baseline characteristics will be summarized for each cohort 
using the statistics of number, mean, median and range for continuous variables and for discrete factors, values 
will be tabulated.    
Safety Analysis: Descriptive statistics will be used in the reporting of adverse events. Adverse events 
will be tabulated and frequencies of events will be determined. All events with a toxicity of Grade 3 or above 
(except as discussed in section 4.2) will be tabulated by event, as well as tabulations for all events (where toxicity 
is defined by the CTCAE, version 4.0). Laboratory analyses (chemistries, hematology, urinalysis, 
serological/immunological analyses, WBC and differential) will consist of measurements of change from 
baseline over time by patient and overall, with plots of actual values compared to normal values for patients by 
dose group. Logarithmic transformations may be applied as necessary. Group means and standard errors will be 
calculated for the various laboratory parameters. Concurrent Illnesses will be listed and examined by univariate 
and multivariable analysis as possible confounders in the treatment response relationship. Concurrent 
medications will also be listed. Effects of previous treatments for cancer will also be examined by univariate and 
multivariable analysis, and any potential related side effects will be analyzed and discussed. 
11.2 Sample Size/Accrual Rate 
Because the number of patients enrolled at each dose level depends on observed toxicities, it is not 
possible to state a definite sample size that will be accrued. The dose escalation plan does allow for a minimum 
of 4 additional patients to be enrolled (if the first two patients enrolled experience dose-limiting toxicity as do 
patients at each of two CRM-defined de-escalated dose levels) and a maximum of 20additional patients to be 
enrolled and treated.  It is expected that the actual enrollment will fall well between these two extremes. 
It is expected that this study will accrue approximately 10-12 patients per year. 
11.3 Stratification Factors 
This study will not stratify patients according to any baseline factors. 
11.4 Analysis of Secondary Endpoints 
11.4.1 Delineation of the local and systemic immune response to M032 administration. 
 
Virus reactivation and shedding will be detected by PCR and culture of serial serum and 
saliva samples. 
 
Immunogenicity of M032 will be evaluated by the use of ELISA to detect HSV antibody 
titers. 
 
Each of these binomial responses will be summarized by frequencies for each cohort and 
overall. 
11.4.2 Gather preliminary information about the potential benefit of M032 in the treatment of patients 
with recurrent malignant gliomas. 
 
The percentage of patients experiencing complete response, partial response, stable disease 
and progressive disease will be reported by cohort and overall based on follow-up 
radiographic imaging. 
 
Changes in clinical disease status and steroid administration will be considered when 
reviewing changes in tumor volumetric size. 
 
All changes in tumor volume will also be analyzed with consideration of any other anti-
tumor cancer therapies (either prior to M032 administration or following M032 failure) and 
the timeframes in which they were administered. 

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 
Quality of life response: A Karnofsky Performance Status score will be recorded pre-
treatment and then measured serially post-treatment for each patient. Time to KPS <60 will 
be measured for each patient by the Kaplan-Meier analytical method.  
12. REFERENCES 
1. 
Martuza, R.L., et al., Experimental therapy of human glioma by means of a genetically 
engineered virus mutant. Science, 1991. 252(5007): p. 854-6. 
2. 
Markert, J.M., et al., Expanded spectrum of viral therapy in the treatment of nervous system 
tumors. J Neurosurg, 1992. 77(4): p. 590-4. 
3. 
Markert, J.M., et al., Reduction and elimination of encephalitis in an experimental glioma 
therapy model with attenuated herpes simplex mutants that retain susceptibility to acyclovir. 
Neurosurgery, 1993. 32(4): p. 597-603. 
4. 
Mineta, T., S.D. Rabkin, and R.L. Martuza, Treatment of malignant gliomas using ganciclovir-
hypersensitive, ribonucleotide reductase-deficient herpes simplex viral mutant. Cancer Res, 
1994. 54(15): p. 3963-6. 
5. 
Chambers, R., et al., Comparison of genetically engineered herpes simplex viruses for the 
treatment of brain tumors in a scid mouse model of human malignant glioma. Proc Natl Acad 
Sci U S A, 1995. 92(5): p. 1411-5. 
6. 
Mineta, T., et al., Attenuated multi-mutated herpes simplex virus-1 for the treatment of 
malignant gliomas. Nat Med, 1995. 1(9): p. 938-43. 
7. 
Markert, J.M., et al., Genetically engineered HSV in the treatment of glioma: a review. Rev Med 
Virol, 2000. 10(1): p. 17-30. 
8. 
Chou, J., et al., Mapping of herpes simplex virus-1 neurovirulence to gamma 134.5, a gene 
nonessential for growth in culture. Science, 1990. 250(4985): p. 1262-6. 
9. 
Andreansky, S., et al., Evaluation of genetically engineered herpes simplex viruses as oncolytic 
agents for human malignant brain tumors. Cancer Res, 1997. 57(8): p. 1502-9. 
10. 
Andreansky, S., et al., Treatment of intracranial gliomas in immunocompetent mice using herpes 
simplex viruses that express murine interleukins. Gene Ther, 1998. 5(1): p. 121-30. 
11. 
MacLean, A.R., et al., Herpes simplex virus type 1 deletion variants 1714 and 1716 pinpoint 
neurovirulence-related sequences in Glasgow strain 17+ between immediate early gene 1 and 
the 'a' sequence. J Gen Virol, 1991. 72 ( Pt 3): p. 631-9. 
12. 
Kesari, S., et al., Therapy of experimental human brain tumors using a neuroattenuated herpes 
simplex virus mutant. Lab Invest, 1995. 73(5): p. 636-48. 
13. 
Yazaki, T., et al., Treatment of human malignant meningiomas by G207, a replication-
competent multimutated herpes simplex virus 1. Cancer Res, 1995. 55(21): p. 4752-6. 
14. 
Markert, J.M., et al., Conditionally replicating herpes simplex virus mutant, G207 for the 
treatment of malignant glioma: results of a phase I trial. Gene Ther, 2000. 7(10): p. 867-74. 
15. 
Markert, J.M., et al., Phase Ib trial of mutant herpes simplex virus G207 inoculated pre-and 
post-tumor resection for recurrent GBM. Mol Ther, 2009. 17(1): p. 199-207. 
16. 
Rampling, R., et al., Toxicity evaluation of replication-competent herpes simplex virus (ICP 34.5 
null mutant 1716) in patients with recurrent malignant glioma. Gene Ther, 2000. 7(10): p. 859-
66. 
17. 
Papanastassiou, V., et al., The potential for efficacy of the modified (ICP 34.5(-)) herpes simplex 
virus HSV1716 following intratumoural injection into human malignant glioma: a proof of 
principle study. Gene Ther, 2002. 9(6): p. 398-406. 
18. 
Harrow, S., et al., HSV1716 injection into the brain adjacent to tumour following surgical 
resection of high-grade glioma: safety data and long-term survival. Gene Ther, 2004. 11(22): 
p. 1648-58. 
19. 
Lagunoff, M. and B. Roizman, The regulation of synthesis and properties of the protein product 
of open reading frame P of the herpes simplex virus 1 genome. J Virol, 1995. 69(6): p. 3615-23. 

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20. 
Parker, J.N., et al., Engineered herpes simplex virus expressing IL-12 in the treatment of 
experimental murine brain tumors. Proc Natl Acad Sci U S A, 2000. 97(5): p. 2208-13. 
21. 
Hellums, E.K., et al., Increased efficacy of an interleukin-12-secreting herpes simplex virus in 
a syngeneic intracranial murine glioma model. Neuro Oncol, 2005. 7(3): p. 213-24. 
22. 
Parker, J.N., et al., Enhanced inhibition of syngeneic murine tumors by combinatorial therapy 
with genetically engineered HSV-1 expressing CCL2 and IL-12. Cancer Gene Ther, 2005. 12(4): 
p. 359-68. 
23. 
Long, Z., et al., Biosafety monitoring of patients receiving intracerebral injections of murine 
retroviral vector producer cells. Hum Gene Ther, 1998. 9(8): p. 1165-72. 
24. 
Levine, A., in Cancer:  Principles and Practice of Oncology, V. DeVita, S. Rosenberg, and S. 
Hellman, Editors. 1989, Lippincott Williams and Wilkins. 
25. 
Chou, J. and B. Roizman, The gamma 1(34.5) gene of herpes simplex virus 1 precludes 
neuroblastoma cells from triggering total shutoff of protein synthesis characteristic of 
programed cell death in neuronal cells. Proc Natl Acad Sci U S A, 1992. 89(8): p. 3266-70. 
26. 
Chua, A.O., et al., Expression cloning of a human IL-12 receptor component. A new member of 
the cytokine receptor superfamily with strong homology to gp130. J Immunol, 1994. 153(1): p. 
128-36. 
27. 
Toda, M., et al., In situ cancer vaccination: an IL-12 defective vector/replication-competent 
herpes simplex virus combination induces local and systemic antitumor activity. J Immunol, 
1998. 160(9): p. 4457-64. 
28. 
Caruso, M., et al., Adenovirus-mediated interleukin-12 gene therapy for metastatic colon 
carcinoma. Proc Natl Acad Sci U S A, 1996. 93(21): p. 11302-6. 
29. 
Bramson, J.L., et al., Direct intratumoral injection of an adenovirus expressing interleukin-12 
induces regression and long-lasting immunity that is associated with highly localized expression 
of interleukin-12. Hum Gene Ther, 1996. 7(16): p. 1995-2002. 
30. 
Wu, C.Y., et al., IL-12 induces the production of IFN-gamma by neonatal human CD4 T cells. 
J Immunol, 1993. 151(4): p. 1938-49. 
31. 
Gately, M.K., et al., Regulation of human lymphocyte proliferation by a heterodimeric cytokine, 
IL-12 (cytotoxic lymphocyte maturation factor). J Immunol, 1991. 147(3): p. 874-82. 
32. 
Manetti, R., et al., Natural killer cell stimulatory factor (interleukin 12 [IL-12]) induces T helper 
type 1 (Th1)-specific immune responses and inhibits the development of IL-4-producing Th cells. 
J Exp Med, 1993. 177(4): p. 1199-204. 
33. 
Voest, E.E., et al., Inhibition of angiogenesis in vivo by interleukin 12. J Natl Cancer Inst, 1995. 
87(8): p. 581-6. 
34. 
Asselin-Paturel, C., et al., Transfer of the murine interleukin-12 gene in vivo by a Semliki Forest 
virus vector induces B16 tumor regression through inhibition of tumor blood vessel formation 
monitored by Doppler ultrasonography. Gene Ther, 1999. 6(4): p. 606-15. 
35. 
Kikuchi, T., et al., Antitumor activity of interleukin 12 against interleukin 2-transduced mouse 
glioma cells. Cancer Lett, 1999. 135(1): p. 47-51. 
36. 
Mikami, T., et al., Antitumor effect of recombinant human lymphotoxin on a tumor line of human 
malignant glioma. Hiroshima J Med Sci, 1989. 38(3): p. 103-7. 
37. 
Whitley, R.J., et al., Vidarabine versus acyclovir therapy in herpes simplex encephalitis. N Engl 
J Med, 1986. 314(3): p. 144-9. 
38. 
Kimberlin DW, Whitley RJ. Viral infections of the newborn infant.  In:  Rudolph's Pediatrics, 
22nd edition.  Rudolph CD, Rudolph AM, Hostetter MK, Lister G, Siegel NJ (eds). McGraw 
Hill Medical, New York, March, 2011. 
39. 
Knipe, D, Whitley, RJ, Roizman, B.  Herpes Simplex Virus.  Fields Virology, in press. 2012 
40.  
Okada H, Weller M, Huang R, Finocchiaro G, Gilbert MR, Wick W, Ellingson BM, Hashimoto 
N, Pollack IF, Brandes AA, Franceschi E, Herold-Mende C, Nayak L, Panigrahy A, Pope WB, 
Prins R, Sampson JH, Wen PY, Reardon DA.  Immunotherapy response assessment in neuro-

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oncology: a report of the RANO working group.  Lancet Oncol. 2015 Nov;16(15):e534-42. doi: 
10.1016/S1470-2045(15)00088-1. PMID:  26545842 
41. 
Fong Y, Kim T, Bhargava A, Schwartz L, Brown K, Brody L, Covey A, Karrasch M, 
Getrajdman G, Mescheder A, Jarnagin W, Kemeny N. A herpes oncolytic virus can be delivered 
via the vasculature to produce biologic changes in human colorectal cancer. Mol Ther. 2009;  
17(2):389-94. PMID: 19018254 
42. 
Markert JM, Cody JJ, Parker JN, Coleman JM, Price KH, Kern ER, Quenelle DC, Lakeman 
AD, Schoeb TR, Palmer CA, Cartner SC, Gillespie GY, Whitley RJ.  Preclinical evaluation of 
a genetically engineered herpes simplex virus expressing interleukin-12.  J Virol. 2012 
86(9):5304-13. 
43. 
Geletneky K, Huesing J, Rommelaere J, Schlehofer JR, Leuchs B, Dahm M, Krebs O, von 
Knebel Doeberitz M, Huber B, Hajda J.   Phase I/IIa study of intratumoral/intracerebral or 
intravenous/intracerebral administration of Parvovirus H-1 (ParvOryx) in patients with 
progressive primary or recurrent glioblastoma multiforme: ParvOryx01 protocol.  BMC 
Cancer. 2012 Mar 21;12:99. doi: 10.1186/1471-2407-12-99.  
44. 
Storer B. Design and analysis of Phase I clinical trials. Biometrics (1989)45:925-937 
45. 
Piantadosi S, Fisher JD, Grossman S.  Practical implementation of a modified continual 
reassessment method for dose-finding trials.  Cancer Chemother Pharmacol. 1998;41(6):429-
36.   
 
 

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APPENDIX A 
 
Expected Adverse Events Associated with M032 and Related International Medical Terminology (IMT) Terms 
 
Category 
Adverse Event 
IMT Preferred Term 
Allergy/Immunology 
Allergic 
reaction/hypersensitivity 
Hypersensitivity NOS 
Blood/Bone Marrow 
Leukocytes 
Leucopenia NOS 
Blood/Bone Marrow 
Hemoglobin 
Haemoglobin decreased 
Blood/Bone Marrow 
Hemolysis 
Hemolysis NOS 
Cardiovascular (General) 
Edema 
Oedema NOS 
Cardiovascular (General) 
Thrombosis/embolism 
Thrombosis NOS 
Constitutional Symptoms 
Fatigue 
Fatigue 
Constitutional Symptoms 
Fever 
Pyrexia 
Constitutional Symptoms 
Weight loss 
Weight decreased 
Gastrointestinal 
Nausea 
Nausea 
Hemorrhage 
Hemorrhage/bleeding without 
grade 3 or 4 thrombocytopenia 
Haemorrhage NOS 
Hepatic 
SGPT (ALT) 
Alanine aminotransferase 
increased 
Infection/Febrile Neutropenia 
Infection without neutropenia 
Infection NOS 
Neurology 
Pyramidal tract dysfunction 
Upper motor neurone lesion 
Neurology 
Speech impairment 
Speech disorder NEC 
Neurology 
Memory loss 
Amnesia NEC 
Neurology 
Confusion 
Confusion 
Neurology 
Mood alteration—depression 
Depression NEC 
Neurology 
Personality/Behavioral 
Personality change 
Neurology 
Neurology—Other (Specify, 
Somnolence) 
Not Available 
Neurology 
Depressed level of 
consciousness 
Depressed level of 
consciousness 
Neurology 
Seizure(s) 
Convulsions NOS 
Neurology 
Neurology—Other (Specify, 
Tumor Progression) 
Not Available 
Pain 
Headache 
Headache NOS 
 
Note: The full list of IMT terms is available on the CTEP home page 
(http://ctep.info.nih.gov/CtepInformatics/IMT.htm ). 
 

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APPENDIX B 
 
Performance Status Criteria 
 
Karnofsky Performance Scale 
Percent 
Description 
100 
Normal, no complaints, no evidence of 
disease. 
90 
Able to carry on normal activity; minor 
signs or symptoms of disease. 
80 
Normal activity with effort; some signs 
or symptoms of disease. 
70 
Cares for self, unable to carry on 
normal activity or to do active work. 
60 
Requires occasional assistance, but is 
able to care for most of his/her needs. 
50 
Requires considerable assistance and 
frequent medical care. 
40 
Disabled, requires special care and 
assistance. 
30 
Severely disabled, hospitalization 
indicated.  Death not imminent. 
20 
Very sick, hospitalization indicated. 
Death not imminent. 
10 
Moribund, fatal processes progressing 
rapidly. 
0 
Dead. 
 

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APPENDIX C 
INSTRUCTION FOR M032 DOSE PREPARATION 
Preparation of M032 dilutions will take place in the hospital pharmacy. 
M032 (NSC 733972) should be prepared for administration following Biosafety Level 2 Guidelines using a laminar 
flow biological safety cabinet, hood, gloves, safety glasses and gown. 
MATERIALS NEEDED FOR PREPARATION OF M032 DOSE FOR SUBJECT ADMINISTRATION  
1. 70% Isopropanol solution  
2. MetriSpray, Cavicide or equivalent disinfectant. 
3. 5 cc Becton-Dickson disposable syringes, Product code  (total of 4 syringes needed) 
4. Tuberculin syringe and 18-23 G needle  
5. Sterile Vector Diluent (10% Glycerol in Phosphate Buffered Saline, pH 7.0-7.4, USP, preservative-free for 
injection, 50 cc bottles, Alanza, Inc.)) cooled to 4°C.   
6. Sterile Gloves 
7. Alcohol Swabs/Wipes 
8. Kim-wipes 
EQUIPMENT NEEDED FOR PREPARATION OF M032 DOSE FOR SUBJECT ADMINISTRATION 
1. Laminar Flow Biological Safety Cabinet (LFBSC) 
2. Rack for vector M032 Stock Vials 
3. Refrigerator 
4. Wet ice or coolant 
METHOD FOR PREPARATION OF M032 DILUTION FOR SUBJECT ADMINISTRATION 
1. Remove all items from the LFBSC, wipe down work surfaces with MetriSpray, Cavicide or equivalent 
disinfectant, followed by 70% isopropanol, allow blower to run for 15 minutes prior to using.   
2. Cool virus diluent (sterile preservative-free 10% Glycerol in Phosphate Buffered Saline, pH 7.0-7.4, USP) to 
approximately 4°C in a refrigerator or on ice. 
3. Thaw the appropriate number of vector stock M032 vial(s) according to Table 1 in Appendix D Dose 
Preparation Guide below, by rubbing gently between gloved hands. Continue until the last ice crystals have 
melted and then place vials on ice.  
4. Note the time that the contents have completely thawed. Do not allow the vials to warm.  
5. Wipe stock M032 vials dry with Kim-Wipes.  Gently mix contents of stock M032 vials by swirling.  
6. Wipe stock M032 vials with alcohol swab and place in a vial rack in the LFBSC to dry. 
7. Dilute M032 according to Table 1. Dose Preparation Guide 
7.1 
Draw up the required amount of sterile preservative-free Phosphate-buffered saline + 10% glycerol 
(Alanza), USP for injection (vector diluent)   
7.2 
Remove the required volume of stock M032 from the glass vials. Add stock M032 to the 
receptacle/vehicle containing the required diluent 
7.3 
Gently invert end over end at least 5 times to mix contents   

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APPENDIX C 
INSTRUCTION FOR M032 DOSE PREPARATION (CONT) 
8. Label administration syringe(s) appropriately and load 6-10 ml diluted M032 into each syringe. Prepare all 
syringes for each of the three hour infusions. Store syringes that will be used for the 3 hour exchanges at 4 C 
until dispensed to the patient.   
8.1 Intra-tumor infusion via catheters: Use a 10 cc syringe containing 6- 10 cc of diluted M032. The 
syringe will be fitted with a luer lok connection to non-compressible tubing connected to each catheter 
so luer-lok syringes are preferred. 
9. Place syringe(s) in sterile syringe holder. Transport the required number of syringes to the patient.  There 
should be 1 syringe for each active catheter to be used.  The remainder of the loaded syringes may remain in 
the Research Pharmacy at 4C until dispensed to the patient at 3 hours or, if a 4° C refrigerator is available, in 
the CRU.    
10. All remaining biological material must be chemically disinfected or autoclaved.  All instruments coming in 
contact with M032 must be autoclaved.  All other items coming in contact with M032 must be incinerated or 
autoclaved.  
11. Waste Disposal responsibility must be managed according to the institutional SOP on disposal of hazardous 
waste.   
 
 
 
 

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APPENDIX D 
INSTRUCTION FOR M032 DOSE PREPARATION 
FOR GMP LOT 0909005 ONLY 
6 hr Infusion Only 
 
Summary: 
 
The following protocol for preparation of different doses of M032 for infusion is provided for informational 
purposes only and describes the methods used in preparing doses at 1 log intervals, beginning at the starting dose of 1 x 
105 PFU delivered in the total volume of 2.4cc.  The Continual Reassessment Method (CRM) will estimate the exact 
dose to be administered to each subsequent subject in 2.4cc.  This calculation will be made once each subject has met 
the 24 day interval and all relevant events have been recorded.  A subject- and dose-specific protocol will be prepared 
and provided to the Trial PIs, the Research Nurse (Study Coordinator) and the Research Pharmacist within a 24 hour 
period to ensure adequate time to review and approve, before that next subject is enrolled in the study.    
 
 

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APPENDIX D 
INSTRUCTION FOR M032 DOSE PREPARATION (CONT) 
FOR GMP LOT 0909005 ONLY 
6 hr Infusion Only 
 
Dose Preparation Guide for Dose Level One: 
 
 
Dose Preparation Guide for Dose Level 2: 
 
 

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APPENDIX D 
INSTRUCTION FOR M032 DOSE PREPARATION (CONT) 
FOR GMP LOT 0909005 ONLY 
6 hr Infusion Only 
 
Dose Preparation Guide for Dose Level 3: 
 
 
Dose Preparation Guide for Dose Level 4: 
 
 

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APPENDIX D 
INSTRUCTION FOR M032 DOSE PREPARATION (CONT) 
FOR GMP LOT 0909005 ONLY 
6 hr Infusion Only 
 
 
Dose Preparation Guide for Dose Level 5: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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APPENDIX E 
M032 SHIPMENT INFORMATION/HANDLING 
 
 
M032 (NSC 733972) Lot L0909005 will be supplied by the NIH RAID program.  M032 shipment requests or 
questions regarding M032 and investigational drug supply should be addressed to: 
 
Anthony Welch 
Program Director 
Biological Resources Branch, DTP 
Division of Cancer Treatment and Diagnosis 
NCI_Frederick 
Building 1052, Room 249 
Frederick, MD  21702 
Phone: 301-846-5691 
Fax: 
301-846-5429 
Email:  welcha@mail.nih.gov 
 
M032 is formulated in Dulbecco’s phosphate buffered saline (D-PBS), 0.4M NaCl and 10% (w/v) glycerol at a 
concentration of 5.8e9 pfu/ml.  The virus product for the study is supplied in 1.0 ml, single use glass vials with a fill 
volume of 0.50 mls. 
M032 will be shipped on dry ice to the Research Pharmacy, at the following address: 
Rachel Burell, Pharm.D.,  
Senior Pharmacist 
UAB Hospital Investigational Drug Service Pharmacy 
North Pavilion, Room 3470 
1802 6th Avenue South  
Birmingham, AL  35249 
Phone: 205-934-7191 
Upon shipment, contents will be checked against the shipping paperwork supplied by the manufacturer and condition on 
arrival documented. Receipt will be verified by faxing completed paperwork to the manufacturer.  Virus will then be 
logged into the M032 Study Drug Inventory Log. 
M032 will be stored frozen at –75 ± 15 C.  Immediately prior to use, virus should be thawed, diluted, and prepared 
according to the instructions provided in Appendix C.  All vials of M032 must be signed out on the Study Drug Inventory 
Log for inventory control. 
Precautions consistent with institutional standard operating procedures (SOPs) for the handling of viruses (Biosafety 
Level 2) should be maintained during the preparation and administration of M032.  Virus preparation should be 
performed in a laminar flow biological safety cabinet using disposable materials.  Study personnel involved in the 
preparation, administration, and disposal of M032 should wear appropriate safety gown, gloves and glasses (refer to 
Appendix C). 
Unused vials of M032 should be returned to the NIH RAID program at the end of the study as instructed.  The clinical 
site will maintain accurate documentation of drug use and the destruction of partially used vials.  The monitor will check 
these records as part of the review of drug accountability. 
 

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APPENDIX F 
Serious Adverse Event Reporting Form 
 
 
We will utilize MedWatch to report any AEs or SAEs, using Form FDA3500A. A copy of the form can be found on 
the following pages. 

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Page 1 of AE reporting form 
 

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Page 2 of AE reporting form 
 

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Page 3 of AE reporting form  
 

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Page 4 of AE reporting form 
 

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Page 5 of AE reporting form 
 

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Page 6 of AE reporting form 
 

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Page 7 of AE reporting form 
 

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Page 8 of AE reporting form 
 

NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
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NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
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NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
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NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
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NCI  Protocol #:  N/A 
Local Protocol #:  UAB-1317 
M032 Amendment 12 
University of Alabama at Birmingham 
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APPENDIX G 
DATA AND SAFETY MONITORING BOARD 
 
The Independent Data and Safety Monitoring Board will utilize the DSMB plan that has been formulated by the UAB 
Comprehensive Cancer Center and is NCI-approved (Section 4.5, page 21).   
Medical Monitor:  
John W. Gnann, Jr., MD 
Professor of Medicine 
Division of Infectious Diseases 
University of South Carolina 
135 Rutledge Avenue, Suite 1207 
MSC 752 
Charleston, SC 29425 
gnann@musc.edu 
 
 
Chairman:   Steven S. Rosenfeld, M.D., Ph.D.Mayo Clinic 
Jacksonville,. FL 
(In process of moving from Cleveland Clinic – new position, contact and address information incomplete at this time.) 
 
Richard L. Hodinka, PhD 
Emeritus Professor, Department of Pathology and Laboratory Medicine 
Perelman School of Medicine at the University of Pennsylvania 
Clinical Professor, Microbiology 
University of South Carolina School of Medicine Greenville 
Room 227, Health Science Administration Building 
701 Grove Road 
Greenville, SC  29605 
hodinka@greenevillemed.sc.edu 
 
Timothy Cripe, MD, PhD 
Professor and Chief of Hematology/Oncology 
Nationwide Childrens Hospital 
700 Childrens Dr. 
Columbus, Ohio  43205 
timothy.cripe@nationwidechildrens.org   
 
Raphael (Ray) Dolin, MD 
Maxwell Finland Professor of Medicine 
Division of Viral Pathogenesis 
Beth Israel Deaconess Medical Center 
330 Brookline Avenue 
Boston MA  02215 
Rdolin@bidmc.harvard.edu  
 
Gretchen Cloud, MS 
 
 
Statistician 
5518 Parkview Circle 
Birmingham Al 35242 
Cloudstat@charter.net  
 
 

 
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INFORMED CONSENT  
 
TITLE OF RESEARCH: 
A Phase 1 Study of M032 (NSC733972), a Genetically 
Engineered HSV-1 Expressing IL-12, in Patients with 
Recurrent/Progressive Glioblastoma Multiforme, Anaplastic 
Astrocytoma, or Gliosarcoma 
 
IRB PROTOCOL #: 
UAB:  IRB-131004002 
 
Vanderbilt:  131589 
 
PRINCIPAL INVESTIGATOR: 
James M. Markert, M.D. 
 
CO-INVESTIGATOR: 
L. Burt Nabors, M.D. 
 
Barton L. Guthrie, M.D. 
Xiaosi Han, MD 
 
SPONSOR: 
NCI Grant 
 
Purpose of the Research 
 
This study is being conducted by Dr. James M. Markert at the University of Alabama at Birmingham. 
The purpose of this study is to establish a safe dose range based on side effects of the study treatment 
being tested, in this case, M032.  All the patients who take part in this study will receive the same type of 
experimental treatment, although some people will receive higher doses than other people and the 
method of administration may vary.  There is no “placebo” in this study.  The total length of time you 
will be followed for this study is 1 year.  When possible, safety data will be collected for up to 15 years 
from your clinical visits with your standard of care practitioner. 
 
Varying doses of M032 will be administered in this study. There will be no more than 6 subjects in each 
dose group. The dose you receive will be determined by the number of subjects given the drug before 
you. We will only increase the dose you receive if the dose given to those who were treated before you 
was determined to be safe.  Anywhere from 8 to 24 patients are expected to take part and be treated in the 
study; the final number will depend on the safety results.  
 
Explanation of Procedures 
 
You are being asked to participate in a research study because you have a malignant brain tumor that has 
not responded to standard treatment and have elected to undergo a tumor biopsy as part of your standard 
medical care.  This document will tell you about the purpose, risks, and benefits of this study.  You 
should provide your consent only after you have received all the necessary information and have had 
enough time to decide whether you wish to participate.  Please feel free to ask any questions before you 
agree to take part in this study. 
 
This study is being conducted to determine how safe and how well-tolerated the experimental study drug, 
"M032" is when administered into the brain where the tumor is located.   “M032” is a genetically 
modified herpes simplex virus or "HSV" (the virus that usually causes cold sores and rarely, a severe 
infection of the brain). It has been known that viruses may kill tumor cells. When tumor cells are mixed 
with certain viruses in the laboratory, the tumor cells die. The (HSV) virus has been modified so that 
tumor cells may be killed when infected by M032. The changes made to the virus (HSV) should help 
prevent the (M032) virus from infecting normal brain tissue.  Extensive testing in both mice and 
monkeys, paid for by the Federal Government, has demonstrated that M032 is not able to cause disease 

 
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when injected directly into the brain.   M032 may also help kill tumor cells by making a protein that 
should help your body’s own immune system to do a better job of fighting off the tumor.  
 
Twelve Owl monkeys with normal brains were injected with M032 at a dose (100 million active viral 
particles) that is equivalent to a human dose of 5 billion active viral particles (150 times the highest dose 
planned in this clinical trial).  Owl monkeys were selected because there were known to be extremely 
sensitive to HSV infection, similar to a newborn human baby.  In toxicity studies with these monkeys, 
one monkey began losing weight, became lethargic but did not show signs of viral encephalitis (infection 
in the brain).  The researchers killed the animal in order to study its brain and all organ systems at the 
microscopic and molecular levels and saw inflammation in the nervous system but were not able to 
confirm that the cause for its deteriorating condition was due to an HSV infection. 
 
 Summary of Study Design:  
 
This study is divided into the following sections, also called phases:  the Screening Phase, the Treatment 
Phase and the Follow-up Phase.  Before you can participate in the study, certain tests, exams and 
procedures will be performed to make sure that you can be in this study.  This is called the Screening 
Phase.  If you qualify for the study you will then enter the Treatment Phase which is the phase where you 
receive the study drug and then enter the follow-up phase. 
 
During this study a total of 13 blood samples and one (1) urine sample will also be collected and tested to 
evaluate the general state of your health, look at your immunity, including HIV status, and test for the 
presence of the herpes virus.  HIV status is checked because patients with an immune deficiency are not 
eligible to participate in this research study.  Special research blood tests to monitor the body's ability to 
send signals to the immune system to target the tumor will also be done.  The blood samples will be 
obtained by inserting a needle into a vein in your arm (venipuncture).  The total amount of blood that will 
be drawn over the course of this study will be approximately 2 cups. 
 
Samples conjunctival (eye) secretions and saliva will be collected a total of 11 times and tested for 
presence of the herpes virus. Saliva will be collected by placing one sterile, cotton tipped swab into the 
mouth and conjunctival (eye) fluid will be collected by gently touching separate sterile cotton tipped 
swabs to the corner of each eye.   
 
 
This study will include having a routine physical examination performed approximately 8 times.  
Nervous system exams and vital signs assessments will be performed approximately 34 times during this 
study.   
You will also have an electrocardiogram (ECG) to check the electrical activity of your heart and a chest 
x-ray.  
 
You will be placed on an anticonvulsant (anti-seizure drug) at the screening visit.  If you are already 
taking an anticonvulsant, you will have a second one added.  The medications will be taken for a 
minimum of 28 days.  
 
A total of up to 8 MRI (magnetic resonance imaging) scans will be performed on you throughout the 
study period to monitor your tumor. If there are signs or symptoms that the tumor has become larger, 
additional MRIs may be performed if your doctor feels they are necessary for your routine care. It is also 
possible that your doctor may need to take a biopsy sample of the brain, as part of your routine care, to 
determine the cause of any increase in signs or symptoms you may be having. If this is the case, your 
doctor will discuss this with you at the time and explain details of the procedure to you.  
 

 
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One head CT scan (a type of imaging procedure using x-rays) will be performed before administration of 
the “M032” to make sure the catheters are in the proper place.   
 
During part of the MRI and CT procedures, a needle will be placed into your vein (intravenous line or 
"IV") and dye will be injected into your vein. This dye helps to give a better picture of the brain tumor 
and surrounding brain.  If you have a known history of allergic reactions to the intravenous MRI contrast 
material used for these assessments or if you have a pacemaker, ferro-magnetic aneurysm clips, metal 
infusion pump, metal or shrapnel fragments in your body or certain types of stents, you will not be able 
to be in this study. 
 
You will have one surgery to insert between one and four small tubes (catheters) into your tumor and the 
next day you will receive all of the assigned dose of the study drug injected directly through these tubes 
into your tumor over a 6 hour period (if needed may extend to up to 12 hours). It is important to 
understand that, if you decide to take part in this trial, your particular clinical situation has determined 
that this would be the most appropriate portion of the trial for you.   Your doctor will explain to you 
precisely why this trial may be appropriate to your situation, if you decide to take part in this study. 
 
You will be admitted to the hospital on the day of the surgical procedure.  You will also be assigned to 
receive a pre-determined dose of M032 and a MRI of the head to identify the exact location of the tumor. 
You will also be given medication into a vein (intravenous) to help you relax and feel sleepy before the 
surgical procedure. The surgical procedure is called stereotactic surgery because the catheters that will be 
used to deliver the drug need to be inserted precisely into the tumor based on the MRI scan and using a 
special frame explained below. You will receive medication to minimize any pain (local anesthesia) 
before the surgery.  This surgery involves using a special removable frame (stereotactic frame) that can 
be temporarily connected to the outside of your skull with small screws. This frame is used to guide the 
tubing used for injecting M032 in a fixed and precise position so that the study drug can be administered 
precisely into the tumor.  For your doctor to be able to do this stereotactic surgery, you will have a 
stereotactic frame temporarily connected to your skull with small screws. Importantly, if the tumor has 
grown significantly since the preceding MRI such that it’s maximum diameter is more than 5.5 cm, the 
frame will be removed and you will not be allowed to receive drug or participate in the study to minimize 
the chance of you having a complication from the virus   
 
Before the study drug is injected, your doctor will take a small sample of tissue (biopsy) to check for 
tumor cells and verify the diagnosis of your type of brain tumor (glioma). A thin, long needle connected 
to the stereotactic frame will be guided to the tumor. This procedure is routinely performed in subjects 
for whom a brain biopsy is needed. If the diagnosis is confirmed, then the surgical procedure will 
proceed to prepare for infusion of the drug.   
 
If the biopsy does not verify that tumor cells are present or your doctor determines that the area 
containing the tumor cannot be injected, M032 will not be administered.  
 
Some of your tumor sample may be sent to The Translational Genomics Research Institute (TGen) for 
additional testing and research. You will not receive any results from this testing. 
 
Should you have an access device (eg., Rickham or Ommaya reservoir) that accesses either the more 
central portion of the tumor/cyst, or the CSF space, fluid will be aspirated as possible (expected, 2-10cc) 
from the catheter for laboratory studies. These specimens will be obtained at either the pre-op clinic visit 
or the day of surgery, as well as up to daily during the inpatient hospitalization (per the surgeon’s 
discretion), and again at post op visit day 10, Day 28, and Months 3,6,9 and 12. 
 

 
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Information for Women of Childbearing Potential, Nursing Mothers, and/or Men Capable of 
Fathering a Child 
 
All women will also have a serum pregnancy test and the test must be negative within 14 days of starting 
study treatment to be allowed to enter this study.  Additional serum pregnancy tests will be performed 
during the study if needed.  Because it is unknown if M032 affects mother's milk or the developing fetus, 
women who are pregnant or breast-feeding will not be allowed to participate in the study.  In addition, 
men and women must have been using an effective method of birth control before receiving the study 
drug. You must also agree to continue to use "barrier" birth control (condoms) during the study and for 
six (6) months following the administration of the study drug. Should you become pregnant while 
participating in this study, you should inform your treating physician immediately. For two weeks after 
receiving M032, you should avoid intimate contact with pregnant women, infants and young children and 
individuals with decreased immunity (ability to fight infection). Subjects should also refrain from 
donating blood during the trial. 
 
Treatment Plan: 
 
If you voluntarily decide to participate in this study, you will be hospitalized for a total of approximately 
4 days.  On the first day, you will have the surgical procedure described above to place up to 4 small 
tubes called “catheters” into various parts of the tumor.  These catheters are slightly larger than the lead 
of a pencil and will be left in place and the surgical wound will be closed with sutures.  Following the 
surgery, you will be monitored closely in the Neuro-intensive Care Unit overnight.  If your condition is 
stable, your doctor may transfer you to a Clinical Research Unit bed and you will have a CT scan to 
make sure the catheters are in the correct place and have not shifted.  If the surgeon determines that the 
catheters are properly placed in your tumor, a small syringe will be attached to each catheter and all of 
the drug, M032, will be slowly pumped into the brain tumor - this is designated as Day 1.  This infusion 
will take about 6 hours (if needed may extend to up to 12 hours). After the infusion is completed, the 
catheters will be removed at your bedside and you will be closely monitored by your physicians and 
nurses for the next day or two.   Your temperature, blood pressure, breathing and heart rate ("vital signs") 
and nervous system checks will be performed frequently after surgery and during the M032 
administration to monitor your progress.  The nervous system checks may include all or some of the 
following: walking, measures of alertness, muscle strength and any changes in movement. 
 
You will be discharged when your doctor feels you are stable enough to leave the hospital.  Total time in 
the hospital will likely be 3-4 days.    
 
You will return to clinic for 8 follow-up visits (Day 10, Day 28, and Months 3, 6, 9, and 12) after your 
surgery.  At months 2, 4, and 5, you will be asked to come to UAB and have blood drawn and 
conjunctival and saliva samples collected and to meet with the study coordinator.  The study coordinator 
will ask you questions about your current medications and any changes in your health since your last 
visit.  You will not see the doctor at these 3 visits. 
 
Study Procedures: 
 
Screening Phase:  (approximately 2 weeks prior to Treatment Phase) 
• Informed consent will be signed before any study-related procedures are performed. 
• You will be asked questions about your medical history, medications you are taking, and how you 
feel. 

 
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• You will have a complete physical examination and have your pulse, blood pressure, respiratory 
rate, temperature, height and weight checked. 
• You will have a neurological (nervous system) examination to check your general nervous system 
function and muscle strength. 
• Samples of blood, conjunctival (eye) secretions, saliva, and urine will be collected at this time for 
testing.  This includes a pregnancy test for women capable of bearing children. 
• An ECG, chest x-ray and MRI will be performed. MRI’s that have been obtained within 30 days 
of pre-study visit may be used at the discretion of the study doctor. 
• You will be started on anticonvulsant medication. If you are already taking an anticonvulsant 
medication you may have a second one added per the discretion of the study doctor. 
• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Day 0 (biopsy and catheter placement)  
• You will be admitted to the hospital for biopsy surgery and if eligible, catheter placement. 
• You will be asked questions about medications you are currently taking and any changes to your 
health since last visit. 
• Vital signs (pulse, blood pressure, respiratory rate and temperature) will be obtained prior to 
surgery.  
• A MRI will be performed to determine site for the biopsy and catheter placement.   
• You will have a neurological (nervous system) examination to check your general nervous system 
function and muscle strength. 
• After the surgery, you will be admitted to the Neurosurgery Intensive Care Unit where your vital 
signs will be monitored frequently and frequent neurological exams will be performed. 
• A CT scan will be performed to make sure the catheters did not move and are still in the proper 
place (may be completed on Day 1). 
• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Day 1 (drug administration): 
• If your condition is stable, your doctor may transfer you to a Clinical Research Unit bed for drug 
infusion and monitoring. 
• A complete physical exam will be performed and your wound will be examined. 
• If not done on Day 0, a CT scan will be performed to make sure the catheters did not move and 
are still in the proper place. 
• Administration of study drug which will take approximately 6 hours (may last up to 12 hours). 
• Your vital signs will be monitored frequently and frequent neurological exams will be performed 
before, during and after drug administration. 
• You will be monitored for any signs of adverse events or reactions to the drug. 
• After completion of the study drug infusion, the catheters will be disconnected from the infusion 
tubing and capped. They will be removed after 6-18 hours. 
• A blood sample will be collected. 
• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Day 2: 
• Nervous system exams and vital signs assessments will be performed. 
• Wound examination 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing. (Acceptable up to 2 days after visit) 
• Rickham/Ommaya reservoir fluid aspiration (if applicable; per the surgeon’s discretion). 

 
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Day 3: 
• Nervous system exams and vital signs assessments will be performed. 
• Wound examination 
• A MRI scan will be performed on you to monitor your tumor. 
• A blood sample will be collected. (Acceptable up to 2 days after visit) 
• You will be discharged if your doctor feels you are stable enough to leave the hospital. 
• Rickham/Ommaya reservoir fluid aspiration (if applicable; per the surgeon’s discretion). 
 
Day 10 ± 3 days: 
• You will be asked questions about medications you are currently taking. 
• You will have a complete physical examination and have your pulse, blood pressure, respiratory 
rate, and temperature checked. 
• You will have a neurological (nervous system) examination to check your general nervous system 
function and muscle strength. 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing. 
• You will be asked about any changes to your health since your last visit. 
• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Day 28 ± 4 days: 
• You will be asked questions about medications you are currently taking. 
• You will have a complete physical examination and have your pulse, blood pressure, respiratory 
rate, temperature and weight checked. 
• You will have a neurological (nervous system) examination to check your general nervous system 
function and muscle strength. 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing.   
• A MRI scan will be performed on you to monitor your tumor. 
• You will be asked about any changes to your health since your last visit. 
• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Month 2: 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing.   
• You will be asked about medications you are taking and about any changes to your health since 
your last visit. 
 
Month 3: 
• You will be asked questions about medications you are currently taking. 
• You will have a complete physical examination and have your pulse, blood pressure, respiratory 
rate, temperature and weight checked. 
• You will have a neurological (nervous system) examination to check your general nervous system 
function and muscle strength. 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing.   
• A MRI scan will be performed on you to monitor your tumor. 
• You will be asked about any changes to your health since your last visit. 

 
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• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Month 4: 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing.   
• You will be asked about medications you are taking and about any changes to your health since 
your last visit. 
 
Month 5: 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing.   
• You will be asked about medications you are taking and about any changes to your health since 
your last visit. 
 
Month 6: 
• You will be asked questions about medications you are currently taking. 
• You will have a complete physical examination and have your pulse, blood pressure, respiratory 
rate, temperature and weight checked. 
• You will have a neurological (nervous system) examination to check your general nervous system 
function and muscle strength. 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing.   
• A MRI scan will be performed on you to monitor your tumor. 
• You will be asked about any changes to your health since your last visit. 
• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Month 9: 
• You will be asked questions about medications you are currently taking. 
• You will have a complete physical examination and have your pulse, blood pressure, respiratory 
rate, temperature and weight checked. 
• You will have a neurological (nervous system) examination to check your general nervous system 
function and muscle strength. 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing.   
• A MRI scan will be performed on you to monitor your tumor. 
• You will be asked about any changes to your health since your last visit. 
• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Month 12: 
• You will be asked questions about medications you are currently taking. 
• You will have a complete physical examination and have your pulse, blood pressure, respiratory 
rate, temperature and weight checked. 
• You will have a neurological (nervous system) examination to check your general nervous system 
function and muscle strength. 
• Samples of blood, conjunctival (eye) secretions and saliva will be collected at this time for 
testing.   
• A MRI scan will be performed on you to monitor your tumor. 
• You will be asked about any changes to your health since your last visit. 

 
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• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Unscheduled Visit: 
• You will be asked questions about medications you are currently taking. 
• Samples of blood, conjunctival (eye) secretions and saliva may be collected at this time for 
testing.   
• You will be asked about any changes to your health since your last visit. 
• Rickham/Ommaya reservoir fluid aspiration (if applicable). 
 
Risks and Discomforts 
Blood Draws: 
There may be some temporary pain, bruising, bleeding or rarely, infection at the site where blood 
samples are drawn from your arm. Although rare, some individuals may become faint during blood 
drawing procedures. These complications are rarely severe.   
 
MRI Scans: 
Magnetic Resonance Imaging (MRI) scans are a painless imaging procedure and are very safe for most 
people. Some discomfort may be experienced since you must lie flat and remain as still as possible in a 
long plastic cylinder for approximately 30-45 minutes. Some people also experience anxiety due to fear 
of being in close spaces. You will be closely observed at all times and can be assisted, if necessary by the 
hospital staff performing the procedure. You may be moved out of the machine at your request or, if you 
are experiencing severe anxiety, you may be given anti-anxiety medication prescribed by your doctor to 
make you less anxious. If you would like, ear plugs are available to you to decrease the knocking noise 
you hear that is made by the machine. Pillows will be placed under your knees to make you comfortable 
and you will be covered with a sheet or blanket to keep you warm, if needed. 
 
A small portion of people may develop brief reactions during administration of the dye used in MRI 
testing, including nausea, headaches, hot flashes and heart palpitations (heart skipping a beat). A smaller 
group of subjects may also be allergic to the dye and may develop a rash, itching, hives, breathing 
difficulties and in extreme cases, death. You will be closely monitored throughout the procedure and if 
an allergic reaction develops, you will be treated promptly. Subjects, who are at risk for injury from MRI 
such as former welders or those with pace makers, aneurysm clips, (metal clips on the wall of a large 
artery), metal infusion pumps, or metal and/or shrapnel fragments, will not be entered into the study.  
Also, there is a risk of "nephrogenic systemic fibrosis" (NSF) which is a fibrosis (scarring and 
contraction) of skin, eye, joints and internal organs due to the contrast agent used for MRIs scans.  While 
uncommon, this can be debilitating. Your kidney function will be determined to be within an acceptable 
range before you will be given a contrast agent so that any risk of developing nephrogenic systemic 
fibrosis due to poor kidney function will be minimized.    
 
CT Scan and Chest x-ray: 
The CT scan procedure is a special x-ray procedure that involves lying flat and as still as possible under a 
scanning machine and typically lasts about 15 minutes. Some subjects may experience an uncomfortable 
warm flushing sensation when the dye is injected. The dye could cause problems with kidney function or 
an allergic reaction resulting in a rash, itching, hives, breathing difficulties and in extreme cases, death. 
You will be closely monitored throughout the procedure and if an allergic reaction develops, you will be 
treated promptly. The amount of radiation you receive from the CT scan is approximately 3 "rem". For 

 
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comparison, the standard limit for occupational radiation exposure is 5 rem/year and the amount from 
natural background radiation in Alabama is about 1 rem/year.   
 
A chest X-ray is a common diagnostic procedure that requires a small dose of radiation.  Although 
radiation is cumulative over a lifetime, small doses from a chest X-ray should not affect your health. 
 
Surgery/Catheter Placement and removal: 
The surgical risks of the stereotactic procedure you will have depend on your condition before the 
surgery and the location and size of your tumor. Risks known to be associated with brain surgery, 
involving catheter placement include: 
• 
Hemorrhage (bleeding) 
• 
Deterioration of nervous system functions such as: 
- 
weakness in the arm and or leg 
- 
loss of sensation over parts of your body 
- 
partial or complete loss of function related to communication, such as speech and 
comprehension 
- 
other functions related to intellectual capacity, such as memory 
• 
Infection and death 
• 
Catheter removal requires placement of a new suture to close the skin and prevent infection 
and this is usually done while you are not under anesthesia. This may cause some mild pain or 
discomfort. Pain medicine will be available to minimize your discomfort during this portion 
of the procedure should you wish it. 
 
The relative risks of these procedures, considering your condition, will be discussed with you by your 
doctor.  You will be assessed closely post-surgery for any signs that the surgery may have any of these 
events. 
 
Brain tumor/tissue biopsy and Infusion of M032: 
Risks of brain tumor/tissue biopsy include: 
• 
Bleeding 
• 
Infection 
• 
Possible deterioration of nervous system function 
• 
Inadvertent inoculation of virus into the ventricles or other non-tumor area 
 
Stereotactic frame placement: 
In order for your doctor to be able to do the surgery, you will have to have a special removable frame 
temporarily attached to your skull with small screws.  Although you will also receive medication to 
minimize any pain (local anesthesia and IV tranquilizer), you may feel some pain right when the frame is 
first attached to your head, and these sites may be sore/tender for several days after the treatment. There 
is a very small risk of a local infection from the site where the screws are placed. 
 
Risks of Herpes Simplex Virus/M032: 
Herpes simplex virus is the virus that usually causes cold sores and rarely, a severe brain infection. It can 
also infect other tissues such as skin and the mucous membranes of the mouth, eyes and urinary tract. 
This research study will involve the injection of a modified herpes simplex virus into the brain. Based on 
all of the laboratory studies done in mice and monkeys, the modification should allow the M032 virus to 
infect and kill tumor cells but not normal brain tissue.  This cannot be assured and the purpose of this 

 
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study is to demonstrate that high doses of the virus can be given without any toxic effects.  The potential 
risks of M032 include, but are not limited to: 
• 
Inflammation of the liver (hepatitis) that could cause death (very rarely) 
• 
Wide-spread viral infection with effects ranging from flu-like symptoms to more severe 
reactions 
• 
Allergic reaction to the virus causing symptoms ranging from itching and hives to severe 
cases, difficulty breathing. 
• 
Infection or inflammation of the brain (encephalitis) or spinal fluid space (meningitis), which 
may cause high fevers, confusion, weakness, problems with alertness or thinking, loss of 
consciousness, neurologic difficulties, seizures and even death. 
 
The virus in this study has been modified to prevent the development of infection of normal brain cells. 
However, if you should develop an infection of the brain, you will be treated with the standard medical 
therapy that is very effective in treating and eliminating this kind of infection.  This therapy to destroy 
the virus uses medications to fight the virus (anti-viral drugs). 
 
Should this anti-viral therapy be needed, it would require that you be hospitalized and receive daily 
intravenous infusion of the anti-viral drug acyclovir or similar drug for 14-21 days. With early treatment, 
anti-viral drugs like acyclovir and others are likely to halt progression of herpes simplex viral infection. 
Your doctor may need to perform tests to help determine if there is such an infection. These might 
include a biopsy of brain tissue or testing of the fluid surrounding the spinal cord and the brain called 
cerebral spinal fluid (CSF). 
 
Your doctor will discuss these procedures with you if it becomes necessary to perform them. The risks of 
a brain biopsy include the possibility of bleeding, infection and in rare cases, deterioration in nervous 
system functioning. If a sample of CSF is needed, a small needle will be placed in the small of the back 
into the space around the spinal cord (lumbar puncture) and a sample of fluid removed. This can result in 
headache and in rare cases, worsening of nervous system functioning. 
 
There is also a risk that the tumor itself may swell as a result of virus injection, causing headache, 
lethargy (sleepiness and tiredness), nausea, vomiting, seizures, neurological deficits or even death. 
Should tumor swelling develop, you will receive treatment with steroids (drugs that decrease 
inflammation and swelling) for as long as it is necessary. If nervous system deficits persist despite steroid 
treatment, there is a chance that your doctor would recommend surgery to remove some or all of the 
swollen tumor and ease the pressure on the surrounding brain. You will be monitored closely throughout 
the trial for signs and symptoms of infection so that you can be treated promptly. 
 
The majority of  people in the United States have already been exposed to herpes simplex virus and may 
have antibodies against the virus. If you do not have antibodies against the virus, it is possible that you 
will develop them after receiving M032. These antibodies help fight infection from the herpes simplex 
virus and are not harmful.  
Risk of Interleukin-12 (IL-12) gene therapy in the brain: 
The risk of interleukin-12 (IL-12) gene therapy in the brain is unknown, but could potentially include 
permanent damage to neurologic function due to inflammation or even the development of an 
autoimmune response in which the body’s infection fighting cells perceive normal brain as infection and 
attack it. This could potentially produce problems similar to Multiple Sclerosis (MS). 
 
Other, unknown effects could also occur as a result of interleukin-12 (IL-12) gene therapy in the brain, 
including but not limited to stroke, bleeding, dangerously low blood pressure, damage to liver or kidney 
function, or even death. 

 
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Risk of accessing Ommaya or Rickham reservoir (only if present) 
The risk of accessing an Ommaya or Rickham reservoir includes pain and bleeding at the site of the 
needle puncture. The procedure will be done under sterile conditions, but there is a remote risk of 
infection. Finally, there is a very small risk of causing bleeding at the catheter tip which could be 
asymptomatic but could potentially result in neurologic deficit. 
 
Unknown Risks 
M032 administration into tumor is new and it is possible that despite our extensive efforts, unforeseen 
problems may occur including the possibility of unknown and possible disabling effects or death. 
 
Benefits 
There may be no direct benefit to you from this study. While it is also possible that this experimental 
treatment may kill some of your tumor cells, there may still be no beneficial effect on the course of your 
illness. Because of your participation in this study, we may learn more about potential ways to treat brain 
tumors. This information may prove useful in the future treatment of patients with brain tumors.  
 
Alternatives 
You are being offered the opportunity to participate in this study after your tumor recurred, despite 
appropriate standard therapies for your disease. Other therapy options have been explained to you, 
including: 
• Gliadel ®, a wafer that releases a chemotherapy agent that is implanted into the area of the 
brain tumor 
• Avastin, a tumor-starving (or anti-angiogenic) therapy that blocks a protein called vascular 
endothelial growth factor, or VEGF. 
• Radiation therapy to the brain 
• Additional surgery, to remove tumor 
• Other Chemotherapy 
 
There are no other standard treatments that have been shown to have significant effects in patients with 
your disease. A variety of experimental studies for the treatment of brain tumors are conducted in 
medical centers around the world, but the benefit of their approaches is yet unknown. In addition, you 
may decline any further treatment for your disease. 
 
If at any time after receiving M032, there are signs or symptoms indicating growth of the tumor, your 
doctor will again discuss alternative therapies that may be of benefit to you. If you are treated with 
alternative therapies after receiving M032, you will still be permitted to continue on the study and be 
monitored for effects of M032. 
 
Confidentiality 
Your medical information will be maintained and processed on a computer for the purposes of quality 
assurance and safety monitoring.  This medical information will be kept confidential, to the extent 
allowed by law, by the study doctor, staff, and authorized institutional personnel and will not be available 
outside of the UAB-Comprehensive Cancer Center unless there is required disclosure by law. However, 
your doctor, the Federal funding agent, the U.S. Food and Drug Administration (FDA), National 
Institutes of Health (NIH) and UAB’s Institutional Review Board (IRB) will be able to inspect your 
medical records and have access to confidential information that identifies you by name. The results of 

 
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the study, including laboratory tests and x-rays may be published for scientific purposes, however your 
identity will not be revealed. 
 
Information relating to this study, including your name, medical record number, date of birth and social 
security number may be shared with the billing offices of UAB and UAB Health System-affiliated 
entities so that claims may be submitted for payment to your insurance company for standard care, non-
study related clinical services and procedures provided to you during the course of this study.  A copy of 
this consent form will be placed in your chart as part of your permanent medical record. 
 
If you are eligible for Medicare, your participation in this research study will be noted in your medical 
record. 
 
A description of this clinical trial will be available on http://www.ClinicalTrials.gov (NCT02062827). 
This Web site will not include information that can identify you. At most, the Web site will include a 
summary of the results. You can search this Web site at any time. 
 
You will be tested for HIV and for hepatitis.  You will be told of your tests results and be offered 
counseling before and after the test.  Positive HIV and hepatitis test results must be report to the local 
health authorities. 
 
Voluntary Participation and  Withdrawal 
Whether or not you take part in this study is your choice.  There will be no penalty if you decide not to be 
in the study. If you decide not to be in the study, you will not lose any benefits you are otherwise owed. 
You are free to withdraw from this research study at any time.  Your choice to leave the study will not 
affect your relationship with this institution.  However, you should return to see the study doctor for 
safety reasons so you can be taken off the study drug and referred for follow-up care. 
 
If you end the study early, you will be asked to have blood tests and the same physical and nervous 
system exams you previously had. It is also possible your doctor will request a MRI at this time. 
 
It is also possible that your doctor may withdraw you from the study without your consent if he/she 
thinks this is in your best interest, or if you are not following the procedures required by the study. If you 
withdraw after the catheter is placed for the first injection of M032, the catheter will be removed and you 
will receive routine follow-up care. Your doctor will discuss any alternatives with you in these cases. 
 
New Findings 
Any significant new findings that develop during the course of the study, which may affect your 
willingness to continue in the research, will be provided to you by Dr. James M. Markert or his staff. 
 
Cost of Participation 
There will be no cost to you for the tests and procedures associated with this research study. All study 
related medications, examinations and medical treatment will be provided at no cost during the twelve-
month study period. This includes charges for the surgical placement of the catheters, injections of 
M032, chest x-ray, EKG, MRIs, CT scan, all clinic visits which are part of the study, and research tests 
performed on blood, saliva, conjunctival secretions and tumor tissue samples.  
 
You and/or your insurance company will be billed for standard of care charges which include the costs 
of the surgical biopsy of your tumor, your NICU one night stay and all costs associated with  these. 
 

 
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If you are eligible for Medicare or other health care reimbursement, certain costs (referred to as "routine 
costs") may be billed to Medicare or your health care (insurance) provider if they are not paid for by the 
sponsor. "Routine costs" include the charges for the medical care and items that you would receive even 
if you were not in this study. If the routine costs are billed to Medicare or your provider, it will be noted 
on the claim that the services were provided in the course of your participation in a clinical trial. 
 
If you are in Medicare Advantage (Medicare managed care plan), you should contact someone at your 
plan before you start a clinical trial. They can provide more information about additional costs you could 
incur from participating in clinical trials. 
  
Payment for Participation in Research 
No compensation is available for taking part in this research study. 
 
Payment for Research-Related Injuries 
The University of Alabama at Birmingham (UAB) has not made any provision for monetary 
compensation in the event of injury resulting from this research study.  In the event of injury, treatment 
will be provided, but is not provided free of charge.  These costs will be billed to you or your insurance 
company in the usual manner. 
 
Questions 
You are invited to ask as many questions regarding this research project as you feel are necessary. If you 
have any questions about the research or a research related injury, Dr. Markert or Dr. Nabors will be glad 
to answer them.  
 
Dr. Markert may be reached by calling 205-934-3411 (pager # 5562).  
Dr. Markert’s mailing address is: 
University of Alabama at Birmingham 
510 20th Street South, 1046 FOT 
Birmingham, AL 35294-3410 
 
 
 
 
 
 
Dr. Nabors may also be reached by calling 205-934-3411(pager # 7394).  
Dr. Nabors’ mailing address is: 
 
University of Alabama at Birmingham 
510 20th Street South, 1020 FOT 
Birmingham, AL 35294-3410 
 
If you have questions about your rights as a research participant, or concerns or complaints about the 
research, you may contact the UAB Office of the IRB, (OIRB) at (205) 934-3789 or toll free at 1-855-
860-3789.  Regular hours for the OIRB are 8:00 a.m. to 5:00 p.m. CT, Monday through Friday. You may 
also call this number in the event the research staff cannot be reached or you wish to talk to someone 
else.  
 
Legal Rights 
You are not waiving any of your legal rights by signing this consent form. 
 

 
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Storage of Specimens for future use 
Since the purpose of this research is to evaluate the potential for the development of M032 for potential 
use in participants with your diagnosis (malignant glioma), it is possible that a commercial product may 
result from this research study.  Blood and any tissue samples obtained from you as part of this research 
study could be part of the development of such a product.  There are no plans to share any profits with 
you or your heirs that may result from such a commercial product. 
 
Remnant (what is left over after clinical tests completed) blood and brain tumor tissue samples collected 
during this study may be stored for future use.  Specimens will be identified by your subject #, date of 
birth, and initials.  The use of these specimens will be restricted to support assessment of M032 and 
treatments for glioma.  No commercial development of assays or cell-lines will be undertaken with these 
specimens.  If at any time, you withdraw your consent from participation in this study, the samples will 
be destroyed. 
 
 
 
Initial your choice below: 
 
_______ 
I agree to allow my samples (blood and brain tumor tissue) to be kept and used for future 
research on brain tumors.  
  
 
 
 
 
 
 
 
 
_______ 
I do not agree to allow my samples (blood and brain tumor tissue) to be kept and used for 
future research.  
 
 
 
 
 
SIGNATURES 
 
 
 
 
 
 
 
 
 
 
 
 
Your signature below indicates that you agree to participate in this study. You will receive a copy of this 
informed consent. 
 
I understand that I will be enrolled in this Phase I Clinical Trial of M032 injected into my tumor 
which requires one surgical procedure and approximately 4 days of hospitalization. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Signature of Participant  
 
 
 
 
 
Date 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Signature of Person Obtaining Consent  
 
 
 
Date 
 
Reviewed by: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Signature of Principal Investigator Reviewing Consent Document 
Date 
 

 
Page 15 of 15 
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University of Alabama at Birmingham 
AUTHORIZATION FOR USE/DISCLOSURE OF PROTECTED HEALTH INFORMATION (PHI) FOR RESEARCH 
 
Participant Name:   
 
 
 
UAB IRB Protocol Number: IRB-131004002 
 
Research Protocol:    A Phase 1 Study of a Genetically 
Engineered HSV-1 Expressing IL-12, in Patients with 
Recurrent/Progressive Glioblastoma Multiforme, 
Anaplastic Astrocytoma, or Gliosarcoma 
Principal Investigator:  James M. Markert, MD 
 
Sponsor:  NCI Grant 
 
What is the purpose of this form?  You are being asked to sign this form so that UAB may use and release your protected 
health information for research.  Participation in research is voluntary.  If you choose to participate in the research, you must 
sign this form so that your protected health information may be used for the research. 
 
Why do the researchers want my protected health information?  The researchers want to use your protected health 
information as part of the research protocol listed above and as described to you in the informed consent. 
 
What protected health information do the researchers want to use?  All medical information, including but not limited to 
information and/or records of any diagnosis or treatment of disease or condition, which may include sexually transmitted 
diseases (e.g., HIV, etc.) or communicable diseases, drug/alcohol dependency, etc.; all personal identifiers, including but not 
limited to your name, social security number, medical record number, date of birth, dates of service, etc.; any past, present, 
and future history, examinations, laboratory results, imaging studies and reports and treatments of whatever kind, including 
but not limited to drug/alcohol treatment, psychiatric/psychological treatment; financial/billing information, including but 
not limited to copies of your medical bills,  and any other information related to or collected for use in the research protocol, 
regardless of whether the information was collected for research or non-research (e.g., treatment) purposes. 
 
Who will disclose, use and/or receive my protected health information?  All Individuals/entities listed in the informed 
consent documents, including but not limited to, the physicians, nurses and staff and others performing services related to 
the research (whether at UAB or elsewhere); other operating units of UAB, HSF, UAB Highlands, Children’s of Alabama, Eye 
Foundation Hospital, and the Jefferson County Department of Health, as necessary for their operations; the IRB and its staff; 
the sponsor of the research and its employees and agents, including any CRO; and any outside regulatory agencies, such as 
the Food and Drug Administration, providing oversight or performing other legal and/or regulatory functions for which 
access to participant information is required; including The Translational Genomics Research Institute (TGen). 
 
How will my protected health information be protected once it is given to others?  Your protected health information that 
is given to the study sponsor will remain private to the extent possible, even though the study sponsor is not required to 
follow the federal privacy laws. However, once your information is given to other organizations that are not required to 
follow federal privacy laws, we cannot assure that the information will remain protected. 
 
How long will this Authorization last?  Your authorization for the uses and disclosures described in this Authorization does 
not have an expiration date. 
 
Can I cancel this Authorization?  You may cancel this Authorization at any time by notifying the Principal Investigator, in 
writing, referencing the research protocol and IRB Protocol Number. If you cancel this Authorization, the study doctor and 
staff will not use any new health information for research. However, researchers may continue to use the protected health 
information that was provided before you cancelled your authorization. 
 
Can I see my protected health information?  You have a right to request to see your protected health information. 
However, to ensure the scientific integrity of the research, you will not be able to review the research information until after 
the research protocol has been completed. 
 
Signature of participant:   
 
 
 
 
 
 
 
Date:  
 
 
or participant's legally authorized representative:  
 
 
 
 
 
Date:  
 
 
Printed Name of participant’s representative:  
 
 
 
 
 
Relationship to the participant: