Phase I Trial of a Measles Virus Derivative Producing CEA (MV-CEA) in Patients With Recurrent Glioblastoma Multiforme (GBM)
Phase: PHASE1
Conditions: Anaplastic Astrocytoma, Anaplastic Oligodendroglioma, Mixed Glioma, Recurrent Glioblastoma
Sponsor: Mayo Clinic



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Table of Contents 
MC0671: Phase I Trial of a Measles Virus Derivative Producing CEA (MV-CEA) in Patients with 
Recurrent Glioblastoma Multiforme (GBM) ................................................................................... 1 
Protocol Resources ........................................................................................................................................ 2 
Table of Contents .......................................................................................................................................... 3 
Schema  ......................................................................................................................................................... 4 
1.0 
Background ......................................................................................................................... 5 
2.0 
Goals ................................................................................................................................... 9 
3.0 
Patient Eligibility ................................................................................................................ 9 
4.0 
Test Schedule .................................................................................................................... 12 
5.0 
Grouping Factor ................................................................................................................ 18 
6.0 
Registration/Randomization Procedures ........................................................................... 18 
7.0 
Protocol Treatment ........................................................................................................... 19 
8.0 
Dose Modification ............................................................................................................ 22 
9.0 
Ancillary Treatment .......................................................................................................... 23 
10.0 
Adverse Event (AE) Reporting and Monitoring ............................................................... 24 
11.0 
Treatment Evaluation ........................................................................................................ 27 
12.0 
Descriptive Factor ............................................................................................................. 28 
13.0 
Treatment/Follow–up Decision at Evaluation of Patient .................................................. 29 
14.0 
Pharmacologic/Correlative ............................................................................................... 30 
15.0 
Drug Information .............................................................................................................. 32 
16.0 
Statistical Considerations and Methodology .................................................................... 33 
17.0 
Pathology Considerations ................................................................................................. 35 
18.0 
Data Collection Procedure ................................................................................................ 36 
19.0 
Budget Considerations ...................................................................................................... 36 
20.0 
References ......................................................................................................................... 38 
 

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Schema 
 
Prior to discussing protocol entry with the patient, call the Randomization Center  
(507-284-2753) for Treatment Arm and dose level and to ensure that a place on the protocol  
is open to the patient. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Patients will be accrued Arm A until MTD is determined for Arm A.  Subsequent patients will then be 
accrued to Arm B.  The treatment arms will NOT be open to accrual simultaneously. 
 
 
 
 
Registration 
 
Arm A 
 
Day 1 Resection 
Day 1 MV-CEA 
Arm B 
 
Day 1 Intratumoral 
Administration 
of MV-CEA 
Day 5 Resection 
MV-CEA 
Observation 

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1.0 
Background 
1.1 Glioblastoma multiforme: Glioblastoma multiforme is the most frequent primary brain 
tumor in adults and accounts for the majority of 18,500 primary brain tumor cases diagnosed 
each year in the United States [Jemal, 2005]. It represents one of the most lethal malignancies 
with a median survival of 12-15 months despite multimodality treatment. Gliomas represent a 
promising target for gene transfer approaches given their limited ability to metastasize, but 
despite promising preclinical data [Ram et al, 1997;Shand et al, 1999;Berger et al, 
1997;Oldfield et al, 1993; Rainov, 2000], significant clinical benefit has not been 
materialized to date. Replicating viruses tested in clinical trials for recurrent gliomas include 
herpes viruses [Markert et al, 2000;Rampling et al, 2000] the conditionally replicating 
adenovirus ONYX-015 [Chiocca et al, 2004] and reovirus. In phase I clinical trials of HSV 
strains and of the conditionally replicating adenovirus ONYX-015 in recurrent glioma 
patients, no dose limiting toxicity has been encountered. In vivo replication of these viruses in 
the patients could not be consistently evaluated, however, pointing to inherent difficulties, 
when testing viral therapies in brain tumor patients [Markert et al, 2000], where assessment 
of viral replication requires repeat brain biopsies, an invasive and potentially risky procedure. 
In all completed trials in recurrent glioma patients, the lack of convincing evidence of 
clinically significant antitumor activity points towards the need to further improve the 
existing viral systems or develop novel, more potent viral agents. They also emphasize the 
need to devise systems that could allow monitoring of viral propagation in the tumor in order 
to accomplish dose optimization and individualization of treatment. 
 
1.2 Measles Virus as a cancer therapy: Measles virus is a negative strand, RNA virus, whose 
genome includes six protein products [Griffin et al, 1996]. Three of these proteins participate 
in the formation of the viral envelope; the H-protein is the surface glycoprotein which 
mediates measles virus attachment to its receptors, the CD46 molecule [Dorig et al, 1993], 
and the SLAM receptor (the latter being predominantly present on activated B and T cells). 
The F-protein is responsible for cell fusion after viral attachment has taken place. During 
natural infection with the measles virus, the virus replicates in susceptible tissues causing a 
very characteristic cytopathic effect, with development of multinucleated giant cells 
(syncytia). Cells infected by measles virus express F and H-proteins on their membranes and 
therefore become highly fusogenic. These cells can cause fusion not only with other infected 
cells but also with uninfected neighboring cells. Although wild type measles virus can lead to 
a potentially serious infectious disease, attenuated strains (vaccine strains) of measles virus 
have an excellent safety record and have resulted in significant decreases in measles 
incidence and mortality worldwide [Cutts et al, 1994]. 
 
1.3 Oncolytic measles virus strains have potent antitumor activity against gliomas: Our 
group has developed a novel antitumor approach to treat glioblastoma multiforme. Based on 
the observation that a combination of two of the measles virus membrane glycoproteins, the F 
and H fusogenic membrane glycoproteins had potent antitumor activity against glioma cell 
lines and xenografts, we hypothesized that replicating attenuated vaccine strain of measles 
virus (MV) the natural carrier of these proteins, could amplify this antitumor potential with an 
excellent safety profile. In contrast to normal cells, tumor cells, including glioma cells, 
overexpress the measles virus receptor CD46 which allows for preferential tumor cell 
targeting [Phuong L et al, 2003; Anderson et al, 2004]. In order to develop a method that 
would allow non-invasive real-time monitoring of viral replication in the tumor and dose 
optimization, we created trackable viruses that express the soluble marker CEA.    
 
The MV-CEA virus was constructed by introducing the extracellular N-terminal domain of 

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human CEA in the measles virus backbone derived from the Edmonston vaccine lineage 
[Peng et al, Nat Med, 2002]. CEA is a 677 amino acid glycoprotein that crosses the blood 
brain barrier and can be detected in the serum, as demonstrated in cases of colon cancer, a 
CEA producing malignancy, when it metastasizes to the brain. CEA is not normally 
expressed in gliomas and therefore, measurable levels of CEA peptide in the serum after viral 
treatment would allow us to monitor in vivo the spread and/or elimination of the virus and 
measure the profile of viral gene expression over time.  
 
Infection of a variety of glioblastoma cell lines, both established and primary cell lines 
derived from Mayo Clinic glioblastoma patients and maintained as subcutaneous xenografts, 
showed significant cytopathic effect consisting of cell to cell fusion and extensive syncytia 
formation, leading to apoptotic cell death [Phuong L et al, 2003]. The antitumor effect of 
MV-CEA was assessed in established subcutaneous U87 glioma xenografts in BALB/c nude 
mice. Animals with established subcutaneous U87 xenografts treated with MV-CEA (total 
dose of 8 x 107 TCID50) had significant tumor regression and prolongation of survival as 
compared to control mice (p<0.001 and p=0.007, respectively). Furthermore, treatment of 
intracranial U87 xenografts with MV-CEA at a total dose of 1.8 x 106 TCID50 led to 
statistically significant  tumor regression and prolongation of survival as compared to control 
animals (p=0.0028 and p=0.02, respectively), with 60% of the animals remaining alive and 
tumor free at 80 days from tumor inoculation [Phuong et al, 2003]. Significant antitumor 
efficacy of measles virus strains was also observed against a variety of other animal glioma 
models, including orthotopic glioblastoma models deriving from glioblastoma primary lines, 
such as GBM39 (p=0.0079) and GBM14 (p=0.037) [Petell and Galanis, unpublished data]. 
 
In all preclinical efficacy studies, the kinetic profile of viral gene expression in mice could 
easily be followed by determining serum levels of the virally encoded marker peptide (CEA). 
CEA correlated well with tumor response and the significant antitumor efficacy of the 
engineered virus was not compromised [Phuong et al, 2003]. 
 
1.4 Preclinical assessment of toxicity in Rhesus macaques and in a transgenic mouse model 
of measles virus pathogenecity: Attenuated vaccine strains of measles virus of the 
Edmonston vaccine lineage from where our viruses also derived have an excellent safety 
record with millions of individuals having been safely vaccinated [Cutts et al, 1994]. 
Nevertheless, in our trial we exploit a different route of administration and the proposed dose 
escalation reaches doses that are higher than what normally is used for measles vaccination 
(103 to 104 TCID50). In consultation with the FDA, the preclinical assessment of toxicity 
was performed in non-human primates, Rhesus macaques and Ifnar CD46ko transgenic mice. 
Rhesus macaques (Macacca Mullata) are naturally susceptible to measles virus infection and 
have been extensively used to study measles virus pathogenesis. CNS inoculation of measles 
virus vaccine strains is the standard method by which the measles virus vaccine lots are tested 
for neurotoxicity. Nevertheless, the cost and complexity of studies in primates significantly 
hampers our ability to perform such studies in large numbers of animals. Therefore, 
identifying an appropriate smaller animal model represented a key element in the design of 
our toxicology studies.  
 
Rodents do not normally express the measles virus receptor CD46 and SLAM, therefore, they 
cannot be used to assess toxicity of oncolytic measles virus strains. Dr. R. Cattaneo from the 
Mayo Clinic Molecular Medicine Program has developed a very appealing transgenic mouse 
model of measles virus pathogenecity by inserting a yeast artificial chromosome containing 
the human CD46 receptor gene and knocking out the INF-- receptor gene. These Ifnarko 
CD46 Ge transgenic mice express human CD46 in a tissue distribution that replicates human 

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CD46 expression and are susceptible to MV Edmonston infection. CD46 distribution in the 
brain of CD46 transgenic mice has been shown to mimic the distribution in humans [Mrkic B 
et al, 1998]. Furthermore, studies of intracerebral inoculation of Ifnarko CD46 GE mice with 
MV Edmonston have demonstrated that this can be suitable animal model for neurotoxicity 
assessment [Mrkic B et al, 1998]. 
 
1.5 Evaluation of toxicity after CNS administration of MV-CEA in Rhesus macaques:Five 
adult measles immune Rhesus macaques were used in this toxicology study. Immune animals 
were chosen in order to mimic the study population in the proposed clinical trial. Two 
macaques received 106 TCID50 on days 1 and 5 to mimic the repeat dose administration 
schedule in our trial, two received 105 TCID50 on days 1 and 5, and one animal received 
vehicle control on the same days. The injections were administered in the frontal lobe of the 
animals, a common location for glioblastoma multiforme using previously established 
stereotactic coordinates [Driesse et al, 1998]. By direct brain weight conversion the highest 
dose was 1.5 x higher than maximum human dose in the proposed trial and 3 x higher the 
maximum dose that the normal brain could be exposed in our study. Animals were monitored 
closely for clinical signs including rash, fever, conjunctivitis, and neurologic symptoms such as 
level of consciousness, motor function, gait, activity level, behavior, posture and facial 
symmetry. No evidence of neurological or other toxicity has been observed to date, at 9 months 
from viral administration. Hematological/biochemical parameters were normal at all time 
points. No CEA elevation was detected pointing to absence of virus propagation. There was no 
evidence of viremia in PBMC by QRT-PCR, except for a single time point in one of the high 
dose animals, and no shedding in buccal mucosa secretions was observed. In addition to lack of 
clinical signs, there was no laboratory evidence of encephalitis on CSF exam performed on day 
9 after administration of the first viral dose. No virus was detected in the CSF by QRT-PCR or 
Vero cell overlay. MRI imaging at 4 months revealed no imaging abnormalities. These results 
support the safety of administration of MV-CEA into primary CNS tumors. 
 
1.6 Toxicity after CNS administration of MV-CEA in Ifnarko CD46 Ge mice:The Ifnarko 
CD46 Ge mouse model is a very sensitive animal model of measles virus neurotoxicity. 
Although human patients are not lacking the interferon-- receptor gene, nevertheless, in 
consultation with FDA we applied this model in order to assess the neurotoxicity of MV-CEA 
in immune mice, thus mimicking the worst case scenario in our patient population. One 
hundred and twenty, 6 to 8 weeks old measles immune Ifnarko CD46 Ge mice were assigned to 
three dose groups, one group receiving vehicle control, the second receiving 104 TCID50 on 
days 1 and 5, and the third receiving 105 TCID50 on days 1 and 5. By direct brain weight 
conversion, this is approximately equivalent to 35 x the maximum human dose in our trial. 
General appearance, activity level, body weight and key hematological/biochemical parameters 
were monitored in all groups of animals, without evidence of abnormality. In order to assess 
toxicity, 30 animals from each group were euthanized on early (days 4 and 8), and intermediate 
(day 26) time points and tissues from multiple organs were subjected to histological analysis. 
Close clinical monitoring revealed no signs of clinical toxicity or neurotoxicity. Histologic 
examination of  
ten vital organs—brain, heart, lung, liver, pancreas, kidneys, spleen, ovary, testes, peritoneum, 
and skeletal muscles revealed no significant difference between vehicle treated animals and 
MV-CEA treated animals. No CEA elevation was observed. An additional group of animals is 
due to be euthanized in January 2006 (day 90), in order to assess late toxicity/persistence. 
 
1.7 Safety data from a phase I trial of intraperitoneal administration of MV-CEA in patients 
with ovarian cancer:MV-CEA is currently undergoing phase I testing in ovarian cancer 
patients. Twelve patients have been treated to date in doses ranging from 103 to 106 TCID50. In 

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this phase I trial, MV-CEA is administered in the peritoneal cavity through a peritoneal port-a-
cath. Repeat administration of the virus for up to six cycles is allowed, if good treatment 
tolerance and no disease progression. Intraperitoneal administration of MV-CEA has been well 
tolerated so far with no dose limiting toxicity having been observed. Furthermore, there was no 
evidence of shedding in urine or oral secretions and no evidence of immunosuppression. No 
significant increase in the titers of anti-measles virus antibody in serum or peritoneal fluid after 
repeat administration has been observed. No patient has developed anti-CEA antibodies.  
 
Viral genomes were detected transiently in the peripheral blood mononuclear cells of two 
patients. Preliminary evidence of biologic activity has been observed with 3 patients having > 
50% reduction in the levels of tumor marker CA-125, and 5 patients having stable disease 
lasting from 2 to 9 months as their best objective response.  
  
1.8 Summary of rationale for study design One of the great challenges in clinical applications of 
gene transfer has been our limited ability to monitor viral propagation and transfer of the 
genetic material of interest to the target tissue and, therefore, individualize the treatment in 
order to achieve optimal safety and efficacy. Thus, we have created a recombinant, attenuated 
Edmonston B vaccine strain of measles virus, MV-CEA, that has been genetically engineered 
to express an inert soluble marker peptide (human carcinoembryonic antigen, CEA), and has 
potent antitumor activity against glioma lines and xenografts. The construction of this virus 
variant (which retains its antitumor effect) allows us to monitor the activity of the virus by 
testing CEA levels in the peripheral blood. 
 
Thus, MV-CEA represents a unique new therapeutic agent for glioblastoma multiforme. We 
have shown that viral propagation correlates with CEA levels. Based in these data, we propose 
to proceed with a phase I trial of intratumoral and resection cavity administration of the 
Edmonston’s strain measles virus, genetically engineered to produce CEA in patients with 
recurrent gliomas. 

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2.0 
Goals 
 
2.1 To assess the safety and toxicity of intratumoral and resection cavity administration of an 
Edmonston’s strain measles virus genetically engineered to produce CEA (MV-CEA) in 
patients with recurrent glioblastoma multiforme 
 
2.2 To determine the maximum tolerated dose (MTD) of MV-CEA. 
 
2.3 To characterize viral gene expression at each dose level as manifested by CEA titers. 
 
2.4 To assess viremia, viral replication, and measles virus shedding/persistence following 
intratumoral administration. 
 
2.5 To assess humoral and cellular immune response to the injected virus. 
 
2.6 To assess in a preliminary fashion antitumor efficacy of this approach.  
 
 
3.0 
Patient Eligibility 
Prior to discussing protocol entry with the patient, call the Randomization Center (507-284-
2753) for dose level and to ensure that a place on the protocol is open to the patient. 
 
3.1 
Treatment 
3.11 
Required Characteristics 
 
3.111 18 years. 
 
3.112 Recurrent Grade 3 or 4 glioma, including astrocytoma, 
oligodendroglioma or mixed glioma with histologic confirmation at 
initial diagnosis or recurrence.   
  
3.113 Candidate for gross total or subtotal resection. 
 
3.114 Adequate hematologic, renal and hepatic function tests as indicated by 
the following laboratory values obtained 14 days prior to registration: 
 
ANC 1500/L 
 
PLT 100,000/L 
 
Total bilirubin 1.5 x upper normal limit (ULN) 
 
AST 2 x ULN 
 
Creatinine 2.0 x ULN 
 
Hgb 9.0 gm/dL 
 
PT and aPTT 1.3 x ULN 
 
3.115 Ability to provide informed consent. 
 
3.116 ECOG performance status (PS) 0, 1 or 2. 
 

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3.117 Anti-measles virus immunity as demonstrated by IgG anti-measles 
antibody levels of  1.1 EU/ml as determined by Enzyme Immunoassay  
 
3.118 Normal serum CEA levels (<3 ng/ml) at the time of registration. 
 
3.119a Willing to provide biologic specimens as required by the protocol. 
 
3.119b Negative serum pregnancy test done 7 days prior to registration (for 
women of childbearing potential only). 
 
3.2 
Contraindications 
 
3.21 
Any of the following:  
 
Pregnant women 
 
Nursing women 
 
Men or women of childbearing potential who are unwilling to 
employ adequate contraception 
 
3.22 
Active infection 5 days prior to registration. 
 
3.23 
History of tuberculosis or history of PPD positivity. 
 
3.24 
Any of the following therapies: 
 
Chemotherapy  4 weeks prior to registration (6 wks for nitrosourea-
based chemotherapy) 
 
Immunotherapy  4 weeks prior to registration 
 
Biologic therapy  4 weeks prior to registration 
 
Bevacizumab  12 weeks prior to registration 
 
Non-cytotoxic antitumor drugs, i.e., small molecule cell cycle 
inhibitors  2 weeks prior to registration 
 
Radiation therapy  6 weeks prior to registration 
 
Any viral or gene therapy prior to registration. 
 
3.25 
Failure to fully recover from acute, reversible effects of prior 
chemotherapy regardless of interval since last treatment. 
 
3.26 
New York Heart Association classification III or IV. 
 
3.27 
Requiring blood product support 
 
3.28 
Inadequate seizure control. 
 
3.29a 
Expected communication between ventricles and resection cavity as a 
result of surgery. 
 
3.29b HIV-positive test result, or history of other immunodeficiency. 
 
3.29c 
History of organ transplantation. 
 
3.29d History of chronic hepatitis B or C. 

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3.29e 
Other concurrent chemotherapy, immunotherapy, radiotherapy or any 
ancillary therapy considered investigational (utilized for a non-FDA-
approved indication and in the context of a research investigation). 
 
3.29f 
Exposure to household contacts 15 months old or household contact 
with known immunodeficiency. 
 
3.29g Allergy to measles vaccine or history of severe reaction to prior measles 
vaccination. 
 

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1. Neurologic examination to be performed by neurologist. 
2. For women of childbearing potential only. Must be done 7 days prior to registration. 
3. Day 3 MRI can be performed earlier at the surgeon’s discretion if clinically indicated. 
4. Only if MRI contraindicated 
5. CEA will also be determined in addition at 6 hrs after viral administration and on Days 2, 3, and 5. 
6. LPA=Lymphoproliferative Assay, IFN- ELISPOT; evaluates cellular immunity against measles virus. It will be performed at the laboratory of Dr Greg 
Poland. 
7. Viral shedding will be assessed by RT-PCR of throat gargle specimens and urine samples prior to treatment, and Days 14 and 28 after tumor resection. 
These will be done in the CRU unless patient is hospitalized. 
8. Testing for viremia will be discontinued after day 28 if two consecutive negative tests are obtained. 
9. If viremia is present on Day 28 repeat testing weekly until negative. 
10. Up to 15 years as per gene therapy trial guidelines. 
11. MRI perfusion sequences to be included. 
12. Tumor measurements not required after the 12 month post-progression follow-up. 
13. Patients should be encouraged to return to Mayo Clinic Rochester for this evaluation, but if unable to return, evaluation can be performed by the local 
physician. 
14. Assessments during Observation may be performed by the local physician, provided no evidence of viremia remains and follow-up 
imaging will be provided to study team at Mayo Clinic in Rochester, MN. 
R Research funded. 
 

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6. LPA=Lymphoproliferative Assay, IFN- ELISPOT; evaluates cellular immunity against measles virus. It will be performed at the laboratory of Dr Greg 
Poland. 
7. Viral shedding will be assessed by RT-PCR of throat gargle specimens and urine samples prior to treatment, and days 3, 14 and 28 after tumor resection. 
These will be done in the CRU unless patient is hospitalized. 
8. Testing for viremia will be discontinued after day 28 if two consecutive negative tests are obtained. 
9. If viremia is present on day 28 repeat testing weekly until negative. 
10. Up to 15 years as per gene therapy trial guidelines. 
11. MRI perfusion sequences to be included. 
12. Tumor measurements not required after the 12 month post-progression follow-up. 
13. Patients should be encouraged to return to Mayo Clinic Rochester for this evaluation, but if unable to return, evaluation can be performed by the local 
physician. 
14. Assessments during observation may be performed by the local physician, provided no evidence of viremia remains and follow-up imaging 
will be provided to study team at Mayo Clinic in Rochester, MN. 
R Research funded. 
 

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Cycle 1 
Observation14 
Tests and Procedures 
≤14 days 
prior to 
registration Day 1 Day 3 Day 4 Day 5 
Day 8 
1 day 
14 days 
after 
tumor 
resection  
3 days 
28 days 
after 
tumor 
resection 
3 days 
q 2 mo 
until 
prog 
3 mo 
after 
prog13 
12 mo after 
prog13 and 
yearly10 
thereafter14 
CT or MRI imaging of catheter 
placement R,15 
 
X 
 
 
 
 
 
 
 
 
 
Tumor resection with viral 
administration 
 
 
 
 
X 
 
 
 
 
 
 
1. Neurologic examination to be performed by neurologist.  NOTE:  Weight and ECOG PS are not needed on days 3, 5, and 8) 
2. For women of childbearing potential only. Must be done 7 days prior to registration. 
3. Day 8 MRI can be performed earlier at the surgeon’s discretion if clinically indicated. 
4. Only if MRI contraindicated. 
5. CEA will also be determined in addition at 6 hrs after viral administration and daily Days 2-9. 
6. LPA=Lymphoproliferative Assay, IFN- ELISPOT; evaluates cellular immunity against measles virus. It will be performed at the laboratory 
of Dr Greg Poland. 
7. Viral shedding will be assessed by RT-PCR of throat gargle specimens and urine samples prior to treatment and Days 3, 5, 8, and on Days 14 
and 28 after tumor  resection. These will be done in the CRU unless patient is hospitalized 
8. Testing for viremia will be discontinued after day 28 if two consecutive negative tests are obtained. 
9. If viremia is present on Day 28, repeat testing weekly until negative. 
10. Up to 15 years as per gene therapy trial guidelines. 
11. MRI perfusion sequences to be included. 
12. Tumor measurements not required after the 12 month post-progression follow-up. 
13. Patients should be encouraged to return to Mayo Clinic Rochester for this evaluation, but if unable to return, evaluation can be performed by 
the local physician. 
14. Assessments during Observation may be performed by the local physician, provided no evidence of viremia remains and follow-up imaging 
will be provided to study team at Mayo Clinic in Rochester, MN. 
15. If indicated 
R. Research funded. 
 

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5.0 
Grouping Factor 
Treatment Group:  Arm A vs Arm B 
6.0 
Registration/Randomization Procedures 
6.1 
Prior to discussing protocol entry with the patient, call the Randomization Center  
for dose level and to ensure that a place on the protocol is open to the 
patient. 
 
6.2 
To register a patient, call 
or 
 a completed eligibility checklist to the 
Randomization Center between 8 a.m. and 5 p.m. central time, Monday through Friday. 
 
6.3 
A signed HHS 310 form must be on file in the Randomization Center before an 
investigator may register any patients. 
 
6.4 
At the time of registration, Randomization Center personnel will verify the following: 
 
IRB approval 
 
Patient eligibility 
 
Existence of a signed consent form 
 
Existence of a signed authorization for use and disclosure of protected health 
information. 
 
In addition, the following will be recorded: 
 
Patient has/has not given permission to store samples for future research of 
cancer; 
 
Patient has/has not given permission to store samples for further research or learn 
about, prevent, or treat other health problems 
 
Patient has/has not given Mayo permission to give their samples to outside 
researchers. 
 
6.5 
Treatment on this protocol must be administered at Mayo Clinic Rochester under the 
supervision of a medical oncologist and neurosurgeon. 
 
6.6 
Treatment cannot begin prior to registration and must begin 14 days after registration. 
 
6.7 
Pretreatment tests must be completed within the guidelines specified on the test schedule. 
 
6.8 
All required baseline symptoms (see Section 10.3) must be documented and graded in the 
medical record. 
 
6.9 
Patients will be admitted to the CRU for outpatient visits to complete the throat gargle 
and urine collection for the viral shedding. The clinical research associate (CRA) should 
inquire with the CRU on availability prior to registering the patient. 
 

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7.0 
Protocol Treatment 
7.1 
Treatment Schedule 
Treatment Arm A After confirmation of diagnosis, a tumor resection will be performed. 
The designated viral dose will be diluted in 1 mL of NS and administered in the resection 
cavity using a 20-gauge blunt tip needle injected 1 to 2 cm in the brain parenchyma in 
multiple (10) injection sites. 
 
Dose Level 
Day 1 
As assigned by 
MCCC 
Randomization 
Center 
En block tumor resection with confirmation of diagnosis + 
Injection of MV-CEA in resection cavity 
 
 
Agent 
Dose Level 
Dose 
Route 
Day 
MV-CEA 
0 
104 TCID50 
RC 
1 
“ 
1* 
105 TCID50 
RC 
1 
“ 
1.5** 
3*105 TCID50 
RC 
1 
“ 
2 
106 TCID50 
RC 
1 
“ 
3 
107 TCID50 
RC 
1 
*starting dose level 
** only if 1/6 DLTs are observed at dose level 1 
RC= Administration in resection cavity 
 
 
If any DLT is observed at dose level 1 and the total number of DLTs in this dose level is <2/6, 
dose will be escalated to the intermediate dose level of 3 x 105 TCID50.   
 
Treatment Arm B  
A silastic ventricular catheter will be placed using stereotactic equipment (Stealth, 
Compass, or Leksell systems). CT or MRI imaging will be employed to secure catheter 
placement, if indicated and the designated dose of MV-CEA will be injected into the 
tumor as a single bolus via the catheter. Based on experience from prior gene transfer 
studies with intratumoral administration of viral agents, we plan to dilute the virus in 1 
mL of normal saline [Lang et al, 2003]. The agent will be administered within a 10-
minute period at 0.1 mL per minute, followed by infusion of 0.2 mL of Normal Saline.   
 
The catheter will be left in place to mark the injection site, it will be secured to the dura, 
and the incision will be closed. The procedure has been previously employed in viral  
 
CNS trials with excellent tolerance [Lang et al, 2003]. The patients will have an open 
craniotomy on day 5, timing corresponding to the expected maximum viral replication 
[Phuong et al, 2003]. At that time an en block tumor resection will be performed by 
circumferentially dissecting the tumor using computer-assisted stereotactic technique, 
making sure not to dislodge the catheter. After resection, the same dose of virus diluted in 
1 mL of NS, will be administered circumferentially around the tumor bed, using a 20-

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gauge blunt tip needle injected 1 to 2 cm into the brain parenchyma in multiple (10) 
injection sites, in an equally spaced pattern. Brain regions defined as potentially 
functioning based on stereotactic guidance will not be injected. 
 
Dose Level 
Day 1 
Day 5 
As assigned by 
MCCC 
Randomization 
Center 
Intratumoral administration 
of MV-CEA  
En block tumor resection + Injection of 
MV-CEA in resection cavity 
 
 
Agent 
Dose Level 
Dose 
Route 
Day 
MV-CEA 
-1 
104 TCID50* 
IT/RC 
1 and 5 
“ 
0 
105 TCID50* 
IT/RC 
1 and 5 
“ 
1* 
106 TCID50* 
IT/RC 
1 and 5 
“ 
2 
107 TCID50* 
IT/RC 
1 and 5 
*starting dose level if MTD for Arm A is 107 TCID50 IT/RC= Intratumoral administration (day 1) and  
administration in resection cavity (day 5) 
 
 
Accrual in Arm B will start after the MTD in Arm A has been established. The MTD from Arm A 
will be used to determine the starting dose for Arm B; Arm B starting dose will be one dose level 
less than the MTD found in Arm A (e.g., if the MTD for Arm A is 107 TCID50 then starting dose 
for Arm B will be 106 TCID50 days 1 and 5) 
 
Note:  As of 10/20/2009 the MTD for Arm A has been established as 107 TCID50. Therefore the 
starting dose level for Arm B is dose level 1, (106 TCID50). 

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7.2 
For this protocol, dose-limiting toxicity (DLT) will be defined as an adverse event 
attributed (definitely, probably, or possibly) to the study treatment and meeting the 
following criteria: 
 
Toxicity 
Definition 
Hematologic 
grade 3 as per NCI Common Terminology Criteria for Adverse Events 
v3.0 except grade 3 ANC lasting 72 hours. 
Nonhematologic 
grade 3 as per NCI Common Terminology Criteria v3.0* 
Neurologic toxicity 
grade 2 as per NCI Common Terminology Criteria v3.0 
Allergic reaction 
The grade 2 allergic reactions asymptomatic bronchospasm and/or 
urticaria, and grade 3 allergic reactions 
Viremia** 
Lasting for 6 weeks from last viral administration 
* Grade 3 nausea, vomiting, or diarrhea will be considered dose-limiting only if patient is receiving 
the maximum supportive care regimen described in the protocol. Alopecia will not be 
considered dose limiting. 
**Viremia is defined as detection of any titer of the virus by RT-PCR in patient’s  
 PBMCs. Detection limit in this assay is 1000 genome copies/g RNA. 
 
7.3 
Both arms will use a cohort of 3 design to determine the MTD of both schedules. The 
MTD from Arm A will be used to determine the starting dose Arm B. Cohorts of 3 
patients will be accrued at a dose level, treated and assessed for toxicity 2 weeks after 
treatment (see exception below for dose level 1, Arm A). A minimum of three and 
maximum of 6 patients will be accrued to each dose level per cohort of 3 design 
characteristics. 
 
7.31 
The first three evaluable patients accrued to dose level 1 on Arm A will be 
enrolled sequentially. Each patient will be treated and followed for three weeks 
to assess for toxicity before the next patient is enrolled and treated.  
 
7.4 
MTD is defined as the dose level below the lowest dose that induces dose-limiting 
toxicity in at least one-third of patients (at least 2 of a maximum of 6 new patients).  
 
7.41 
Three patients will be treated at a given dose level and observed for at least 14 
days post-resection to assess toxicity. 
 
7.42 
If dose-limiting toxicity (DLT) is not seen in any of the first 3 patients, 3 new 
patients will be accrued and treated at the next higher dose level. If DLT is seen 
in 2 or 3 of 3 patients treated at a given dose level, then the next 3 patients will be 
treated at the next lower dose level. 
 
7.43 
If DLT is seen in 1 of 3 patients treated at a given dose level, up to 3 additional 
patients will be enrolled and treated at the same dose level. If DLT is seen in at 
least one of these additional three patients (2 of 6), the MTD will have been 
exceeded and further accrual will cease to this cohort. If dose-limiting toxicity 
(DLT) is not seen in any of the three additional patients, 3 new patients will be 
accrued and treated at the next higher dose level. 
 
7.44 
After enrolling 6 patients on a specific dose level, if DLT is observed in at least 2 
of 6 patients, then the MTD will have been exceeded and defined as the previous 
dose unless only 3 patients were treated at the lower dose level. In that case, 3 

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additional patients will be treated at this lower dose level such that a total of 6 
patients are treated at the MTD to more fully assess the toxicities associated with 
the MTD. 
 
7.45 
If one DLT is observed out of 6 patients accrued to dose level 1 on Arm A, we 
will accrue patients to a middle dose level of 3*105. 
 
7.5 
Once the MTD is determined per Section 7.4 for Arm B, an additional 4 patients (for a 
total of 10) will be treated at the Arm B MTD to further assess toxicity and translational 
endpoints (see section 14). 
 
7.6 
Dose de-escalation: If dose-limiting toxicity is seen 2 or more patients at dose level 1 for 
either arm, 3 patients will be entered at dose level 0. 
 
7.61 
If no DLT is observed, the MTD will have been determined. If accruing to Arm 
B, additional patients will be enrolled at dose level 0 as per Section 7.44 to better 
assess toxicity and translational endpoints. If DLT is seen in 1 of 3 patients, up to 
3 additional patients will be enrolled to dose level 0 as per Section 7.43.  If DLTs 
are observed in 2 of 6 patients treated at dose level 0 in Arm A, no further doses  
will be evaluated the study will close to accrual. 
 
7.7 
If a patient fails to complete the initial course of therapy for reasons other than toxicity, 
the patient will be regarded as treatment intolerant and an additional patient will be 
treated at the current dose level; however, all toxicity information will be utilized in the 
analysis. For these instances, a specific notation will be made for review by the Cancer 
Center Clinical Research Administrative Subcommittee (CCCRAS). 
 
7.71 
Patients will be removed from the study if any of the following occur; a) patient 
refuses to continue, regardless of reason; b) the principal investigator deems it in 
the patient’s best interest; c) the patient is unable to continue follow-up; d) a 
major treatment violation occurs; e) the patient experiences DLT, a serious 
illness, measles or progressive disease; f) the study is terminated. 
 
7.72 
If patient develops grade  3 neurologic toxicity during the follow-up phase that 
is thought to be possibly, probably or definitely related to the study agent, study 
accrual will be temporarily held until the data are discussed with the FDA. 
 
7.73 
Accrual for this study will cease permanently if: a) any patient experiences a 
grade 5 treatment-related toxicity; b) study accrual is completed as per section 
16.2. 
 
 
8.0 
Dose Modification 
No dose modification is allowed. Patients who experience DLT in Arm B after Day 1 treatment 
will be taken off treatment.   
 

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9.0 
Ancillary Treatment 
 
9.1 
Patients should receive full supportive care while on this study. This includes blood 
product support, anticonvulsants, perioperative steroids, antibiotic treatment and 
treatment of other newly diagnosed or concurrent medical conditions. All blood products 
and concomitant medications such as antidiarrheals, analgesics, anti-emetics received 
from the first administration of study agent until 28 days post-resection are to be recorded 
in the medical record. 
 
Acceptable treatment options for nausea/vomiting include: 
 
ondansetron 4-8 mg po q 8 hours prn or 4 mg IV q 8 hours prn;  
 
granisetron 1 mg po q 12 hours prn or 10 mcg/kg IV q 12 hours prn;  
 
prochlorperazine 5-10 mg po q 6-8 hours prn, 25 mg rectally bid prn, or 2.5-5 mg IV 
by slow infusion (5 mg/min) q 6 hours prn;  
 
Lorazepam 0.5-2 mg po q 4-6 hours prn; 
 
Aprepitant 125 mg po day 1, 80 mg po on days 2 and 3; 
 
Palonosetron 0.25 mg IV (one time dose).   
 
Acceptable treatment options for diarrhea include: loperamide 4 mg po after first 
diarrheal bowel movement and 2 mg po after each subsequent one up to 16 mg a day and 
atropine sulfate/diphenoxylate hydrochloride (Lomotil) 2 tab po q 6 hours prn or 10 ml q 
6 hours prn.   
 
Acceptable treatment options for fever include acetaminophen 500 mg po 1-2 tabs q 4-6 
hours prn up to 4 grams a day. 
 
9.2 
Diagnosis of measles in this trial is based on the CDC definition of clinical measles and 
includes: 
 
a generalized rash lasting 3 days, and 
 
temperature 38.3C (101F), and 
 
cough, coryza, and conjunctivitis. 
 
9.3 
Since measles virus immunity is required for employment at Mayo, administration of the 
agent is not expected to result in any risk for nursing or ancillary staff. 
 
9.4 
Should a patient develop measles, treatment with intravenous immune globulin will be 
administered 400 mg/kg/d for 3-5 days. Patients who develop measles will be removed 
from treatment and followed per observation schedule in Section 4.0 for up to 15 years 
for evidence of persistent toxicity. 
 
9.5 
Patients participating in Mayo Clinic Cancer Center Phase I Program clinical trials are 
not to be considered for enrollment in any other study involving a pharmacologic agent 
(drugs, biologics, immunotherapy approaches, gene therapy) whether for symptom 
control or therapeutic intent. 
 

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10.0 
Adverse Event (AE) Reporting and Monitoring 
10.1 
This study will utilize the Common Terminology Criteria for Adverse Events (CTCAE) 
v3.0 for adverse event monitoring and reporting. The CTC v3.0 can be accessed via the 
CTEP home page: http://ctep.cancer.gov/. All appropriate treatment areas should have 
access to a copy of the CTCAE v3.0. 
 
10.11 Adverse event monitoring and reporting is a routine part of every clinical trial. 
First, identify and grade the severity of the event using the CTCAE. Next, 
determine whether the event is expected or unexpected (refer to Sections 10.12 
and 15.0) and if the adverse event is related to the medical treatment or procedure 
(see Section 10.13). With this information, determine whether an adverse event 
should be reported as an expedited report (see Section 10.2) in addition to the 
routinely reported clinical data (see Sections 10.31 and 18.0). 
 
Expedited adverse event reporting requires submission of a written report, but 
may also involve telephone notifications. Telephone and written reports are to be 
completed within the timeframes specified in Section 10.2. All expedited adverse 
event reports should also be submitted to the local Institutional Review Board 
(IRB).   
 
10.12 Expected vs. Unexpected 
 
The determination of whether an AE is expected is based on agent-specific 
adverse event information provided in Section 15.0 of the protocol. 
 
Unexpected AEs are those not listed in the agent-specific adverse event 
information provided in Section 15.0 of the protocol. 
 
10.13 Assessment of Attribution 
 
When assessing whether an adverse event is related to a medical treatment or 
procedure, the following attribution categories are utilized: 
 
Definite - The adverse event is clearly related to the investigational agent(s). 
Probable - The adverse event is likely related to the investigational agent(s). 
Possible - The adverse event may be related to the investigational agent(s). 
Unlikely - The adverse event is doubtfully related to the investigational agent(s). 
Unrelated - The adverse event is clearly NOT related to the investigational 
agent(s) 

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10.2 
Expedited Adverse Event Reporting Requirements  
Phase I, II and III Studies (Investigational) 
 
Grade 4 or 51 
Unexpected 
with 
Attribution of Possible, 
Probable, or Definite 
Other Grade 4 or 5 
or 
Any hospitalization 
during treatment6 
Secondary 
AML/MDS2 
Notify the Cancer Center SAE 
Coordinator 3 within 24 hours 
X 
 
 
Submit written report within 5 working 
days4, 7 
X 
 
 
NCI/CTEP Secondary AML/MDS Report 
Form within 15 working days5, 7 
 
 
X 
Submit Grade 4 or 5 Non-AER Reportable 
Events/Hospitalization Form within 5 
working days.6, 7 
 
X6 
 
Submit Serious Adverse Event Reporting 
Form for Human Gene Transfer Clinical 
Studies 3, 7, 8 
X 
 
 
1.  Includes all deaths within 30 days of the last dose of investigational agent regardless of attribution or 
any death attributed to the agent(s) (possible, probable, or definite) regardless of timeframe.  
2.  Reporting for this AE required during or after treatment. 
3. Notify the Cancer Center SAE Coordinator 
Mayo Clinic - Rochester) by telephone
 
 and/or submit a written event summary via fax to 
. The Cancer Center SAE 
Coordinator will subsequently notify the Cancer Center IND Coordinator (see Footnote 7). 
4.  Use Adverse Event Expedited Report – Single Agent or Multiple Agents report form. Submit to the 
Cancer Center SAE Coordinator for further processing (see Footnote 7) and to the Cancer Center 
CRO Safety inbox for IRB reporting. 
5.  Submit per form-specified instructions and provide copy to the Cancer Center CRO Safety inbox for 
IRB reporting and the Cancer Center SAE Coordinator for further processing (see Footnote 7).  
6.  In addition to standard reporting mechanism for this type of event, submit information to the Cancer 
Center CRO Safety inbox for IRB reporting and to the Cancer Center SAE Coordinator for further 
processing (see footnote 7). If Adverse Event Expedited Report – Single Agent or Multiple Agents 
report form was completed, this form does not need to be completed. 
7.  The SAE Coordinator will route these reports to the Cancer Center IND Coordinator who, in 
consultation with the IND holder, will report to the Food and Drug Administration (FDA) as 
warranted by the event and required by U.S. regulations.  
8.  This reporting is in addition to all other reporting.  Submit form to the Office of Recombinant DNA 
Activities, NIH, MSC 7010, 600 Executive Boulevard, Suite 302, Bethesda MD 20892-7010; Phone 
.  Submit copies of the report to the Cancer Center CRO Safety inbox for IRB 
reporting and to the Cancer Center SAE Coordinator for further processing (see Footnote 7).   
 

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10.3 
Adverse events to be graded at each evaluation and pretreatment symptoms/conditions to 
be evaluated at baseline per Common Terminology Criteria for Adverse Events (CTCAE) 
v3.0 grading unless otherwise stated in the table below: 
 
Category 
Adverse 
events/Symptoms 
Baseline 
Each evaluation 
Constitutional 
Symptoms 
Fever (in the absence of 
neutropenia, where 
neutropenia is defined 
as ANC <1.0 x 109/L) 
X 
X 
Rigors/Chills 
X 
X 
Fatigue 
X 
X 
Dermatology/Skin 
Rash/desquamation 
X 
X 
Gastrointestinal 
# of stools per day 
X 
 
Diarrhea 
 
X 
Pain 
Pain – Head/headache 
X 
X 
Neurology 
Neuropathy: motor 
X 
X 
Neuropathy: sensory 
X 
X 
Ataxia (incoordination) 
X 
X 
Seizure 
X 
X 
Speech impairment 
X 
X 
Confusion 
X 
X 
 
 
10.31 Submit via appropriate MCCC Case Report forms (i.e., paper or electronic, as 
applicable) the following AEs experienced by a patient and not specified in 
Section 10.3 
 
10.311 
Grade 1 and 2 AEs deemed possibly, probably, or definitely related to 
the study treatment or procedure. 
 
10.312 
Grade 3 and 4 AEs regardless of attribution to the study treatment or 
procedure 
 
10.313 
Grade 5 AEs (Deaths) 
 
10.3131 
Any death within 30 days of the patient’s last study 
treatment or procedure regardless of attribution to the study 
treatment or procedure 
 
10.3132 
Any death more than 30 days after the patient’s last study 
treatment or procedure that is felt to be at least possibly 
treatment related must also be submitted as a Grade 5 AE, 
with a CTCAE type and attribution assigned. 

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11.0 
Treatment Evaluation 
For the purposes of this study, patients should be re-evaluated at 28 days after tumor resection 
and every 2 months thereafter until progression. If objective response is observed, a confirmatory 
scan is required at 4 weeks after the objective response. 
 
11.1 
Response criteria:  The neurologic examination and the MRI and/or CT at each 
evaluation will be scored as follows: 
 
11.11 
NEURO EXAM STATUS (compared to pre-Rx exam) 
 Better: 
must be on stable or decreasing dose of steroids. 
 Same: 
failure to qualify for better or worse. 
Worse: 
includes patients requiring increasing steroid dose to 
remain stable. 
 
 
11.12 MRI AND/OR CT ASSESSMENT (compared to pretreatment scan for 
bidimensionally measurable disease): 
 
 CR = 
total disappearance of all tumor with patient off corticosteroids or 
only on adrenal replacement maintenance. 
 
PR= 
50% reduction in product of perpendicular diameters of contrast 
enhancement or mass with no new lesions with the patient being 
on stable or decreased steroid dose. 
STAB = 
failure to qualify for CR, PR, REGR, or PROG. 
PROG = 
>25% increase in product of perpendicular diameters of contrast 
enhancement or mass or appearance of new lesions. 
 
 
11.13 MRI AND/OR CT ASSESSMENT (compared to pretreatment scan) for 
evaluable disease (i.e., contrast enhancing mass on MRI and/or CT that is not 
bidimensionally measurable but clearly evaluable for response to therapy. 
 
 CR = 
total disappearance of all tumor with patient off corticosteroids or 
only on adrenal replacement maintenance. 
REGR = 
unequivocal reduction in size of contrast-enhancement or decrease 
in mass effect as agreed upon independently by primary physician 
and quality control physicians; no new lesions.  Patient should be 
on stable or decreased steroid dose. 
STAB = 
failure to qualify for CR, PR, or PROG. 
PROG = 
unequivocal increase in size of contrast enhancement or increase in 
mass effect as agreed upon independently by primary physician 
and quality control physicians:  appearance of new lesions. 
 
 
 

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11.14 Determination of Objective Response Status 
 
NEURO 
MRI and/or CT Status 
STATUS 
CR 
PR 
REGR 
STAB 
PROG 
Better 
 
 
 
 
UNKN* 
 
CR 
PR 
REGR 
STAB 
 
Same 
 
 
 
 
 
 
 
 
 
 
PROG 
Worse 
 
 
UNKN* 
 
 
*Re-image at next evaluation as per protocol and assess as per table below: 
 
 
NEURO 
MRI and/or CT Status 
STATUS 
CR 
PR 
REGR 
STAB 
PROG 
Better 
 
 
 
 
 
 
CR 
PR 
REGR 
STAB 
 
Same 
 
 
 
 
PROG 
 
 
 
 
 
 
Worse 
 
 
 
 
 
 
 
11.15 Survival and Time to Progression Definitions 
 
11.151 
Progression-free survival is defined as the length of time from date of 
registration to a) date of progression or death due to any cause or b) 
last follow-up. 
 
11.152 
Overall survival is defined as the length of time from date of 
registration to a) death due to any cause or b) last follow-up. 
 
 
12.0 
Descriptive Factor 
12.1 
Age (years):  <40 vs. 40-60 vs. >60. 
 
12.2 
Enzyme-inducing anticonvulsant use at study entry:  Yes vs. no. 
 
12.3 
ECOG PS:  0 vs. 1 vs. 2. 
 
12.4 
Prior nitrosoureas:  Yes vs. no. 
 
12.5 
Interval since end of RT (months). 
 
12.6 
Corticosteroid therapy at study entry:  Yes vs. no. 
 
12.7 
Extent of primary resection:  None vs. biopsy vs. subtotal resection vs. gross total 
resection. 
 
12.8 
Extent of resection at recurrence: None vs. biopsy vs. subtotal resection vs. gross total 
resection. 

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12.9a 
Family history of brain tumor:  Yes vs. no. 
If yes, check all that apply: 
_____ Father 
_____ Mother 
_____ Brother/Sister 
_____ Child 
_____ Other (list:___________________________________) 
 
12.9b Primary indicator:  Measurable disease vs. evaluable disease. 
 
12.9c 
Number of prior chemotherapy regimens for progressive/recurrent disease. 
 
 
13.0 
Treatment/Follow–up Decision at Evaluation of Patient 
13.1 
After treatment completion patients will be followed every 2 mos until progression and 
subsequently as per section 4.0. 
13.2 
Patients who have been taken off treatment because of unacceptable toxicity will be 
followed as per observation schedule (Section 4.0).   
 

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14.0 
Pharmacologic/Correlative  
14.1 
Tumor Tissue Analysis 
 
14.11 Arm B: 
Tissue samples from previous surgery or biopsies will be obtained. CD46 
receptor levels will be determined by IHC. The treated tumor will be resected on 
day 5. H&E stains on the resected tumor and surrounding normal brain will be 
performed to assess syncytia formation and cytopathic effect and compared with 
pretreatment tissue sample. If an inflammatory infiltrate is observed it will be 
further characterized by IHC (CD68, CD45, CD3, CD20 stains). The resected 
tumor and surrounding normal brain tissue will be examined by IHC for 
expression of the measles virus N protein and by in situ hybridization and Vero 
cell overlay for the presence of replicating virus. Baseline pre-treatment tissue 
will be tested in comparison. Viral gene expression and replication will be 
characterized in relationship to the distance from the injection site (marked by the 
catheter). Because measles virus induced cell death is mediated by apoptosis, a 
TUNEL assay will also be performed using the  
 
Dead End Colorimetric TUNEL System (Promega). Correlative analysis to assess 
the presence of the virus in tissue and the TUNEL assay will be performed in Dr. 
Galanis’ laboratory using established methodology. 
 
14.12 Arm A: 
The resected tumor will be examined by H & E. If immune infiltrate is present, it  
will be characterized by IHC as described in 14.1. CD46 levels will be 
determined by IHC. 
 
14.2 
Assessment of immune competence: This will be performed by assessment of 
immunoglobulin levels, CD4 and CD8 counts, and CD4/CD8 ratio prior to treatment, and 
at 28 days after tumor resection. 
 
14.3 
Assessment of viremia and viral shedding.  Testing will be performed in Dr. Galanis’ 
laboratory.   
Patient’s peripheral blood mononuclear cells will be monitored for the presence of 
measles virus genome. This will be performed by quantitative RT-PCR for the measles 
virus N mRNA to be performed as follows: a) Arm B: prior to treatment and on days 3, 5, 
8. Also on days 14 and 28 after resection and every 2 months thereafter until progression. 
Subsequent testing as per section 4.0. Testing for viremia will be discontinued after day 
28 if two consecutive negative tests are obtained. b) Arm A; prior to treatment, on days 3, 
14 and 28 after resection and every 2 months thereafter until progression. Subsequent 
testing as per section 4.0. Testing for viremia will be discontinued after day 28 if two 
consecutive negative tests are obtained.  
 
Viral shedding will be assessed by RT-PCR of throat gargle specimen(s) and urine 
samples as follows: Arm B; baseline, on days 3, 8, also on days 14 and 28 after resection. 
Arm A; baseline, on days 3, 14 and 28 after tumor resection. Patients will have the gargle 
and urine specimens done in the CRU as an outpatient visit when patient is not 
hospitalized. Paula Zollman will be the lab contact, located in Guggenheim 1802, phone 
6-0386.   
 

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If a patient is found to be shedding the virus in urine or throat gargle specimens(s), family 
members who don’t have documentation of immunity, will be offered testing to assess 
anti-measles virus immunity by Enzyme Immunoassay (Diamedix, see section 3.18). 
Measles vaccination will be offered to seronegative individuals, as per standard clinical 
practice. 
 
14.4 
Assessment of peripheral immune response to viral administration: measles virus 
specific immunity will be measured by means of (a) measuring anti-measles virus 
specific antibodies (IgG) at baseline, at 28 days after tumor resection and every 2 months 
until progression, and (b) lymphoproliferative assay and IFN- ELISPOT performed at 
baseline and at 28 days after tumor resection.  This testing will be performed in Dr. Greg 
Poland’s laboratory. 
 
14.5 
CEA monitoring as in section 4.0.  The assay will be performed at the Mayo Clinic 
Central Clinical Lab using the Bayer Advia Centaur Immunoassay system (Bayer, 
Tarrytown, NY). 
 
 
 
 

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15.0 
Drug Information 
15.1 
MV-CEA (CEA-Measles Virus – Edmonston Strain) 
15.11 Preparation and storage:  MV-CEA will be prepared at the Vector Production 
Core of the Molecular Medicine Program and stored at -80C. The virus will be 
thawed and mixed with NS immediately prior to administration. 
 
15.12 Known potential toxicities: 
 
15.121 The most common adverse effect noted is burning and stinging at the site 
of injection; however, this is not applicable in this study given CNS 
administration. Occasionally, moderate fever 38.3-39.4C have been 
noted in the month after vaccination, usually within 5-12 days after 
injection.  Rash, which is usually minimal, has been noted. Less 
commonly, high fever over 39.4C or mild lymphadenopathy have been 
reported.  
 
15.122 
Occasional reactions: 
 
Moderate to high fever lasting 1-2 days, starting within a week or 
two of the vaccination 
 
A rash, lasting 1-2 days 
 
Cough and rhinitis 
 
Erythema multiforme (skin rash) 
 
15.123 
Unexpected and rare reactions associated with measles vaccines: 
 Allergic reactions to the vaccine including anaphylaxis  
 Reactions at the injection site such as wheal, flare or urticaria 
 Thrombocytopenia 
 Diarrhea 
 
15.13 Nursing guidelines 
Due to the very early investigational nature of this drug, no nursing guidelines 
are known. Please monitor patients closely and follow Section 10.0. More than 
90% of the U.S. population has measles virus immunity as a result of natural 
infection or immunization. Immune status for measles virus is mandatory for 
Mayo Medical Center personnel. Therefore administration of the agent is not 
expected to result in any risk for nursing or ancillary staff. 
 
 
 
 

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16.0 
Statistical Considerations and Methodology  
16.1 
Study design 
This is a single arm, Phase I study designed to determine the MTD and toxicity of 
attenuated MV-CEA virus in the treatment of recurrent high grade gliomas.   
 
16.11 Sample size:  This phase I study may involve a minimum of 21 patients [12 Arm 
A patients (3 each at dose levels 1 and 2 and 6 at dose level 3) and 9 Arm B 
patients (3 at level 1 and 6 at level 2)] and a maximum of 36 patients [24 Arm A 
patients (6 at each of dose levels 1,1.5,2,3) and 12 arm B patients (6 at dose 
levels 1 and 2)] but would likely require only 24 patients[12 Arm A patients (3 at 
dose levels 1 and 2 and 6 at 3) and 12 Arm B patients (6 at dose levels 1 and 2)]. 
An additional 4 patients will be accrued at the Arm B MTD, for a total of 10 
patients at the MTD, in order 
 
 to further study toxicity and correlative endpoints; thus the study may enroll a 
maximum of 40 patients with an expected number of 28 patients.  
 
16.12 MTD Determination:  MTD is defined as the dose level below the lowest dose 
that induces dose-limiting toxicity in at least one-third of patients (at least 2 of a 
maximum of 6 new patients) or the highest dose level for Arm B (2 x 107 
TCID50), if  1 of 6 patients experience DLT.  See Section 7.4 for the algorithm 
on how the MTD will be determined.   
 
16.13 Accrual and Study Duration:  Accrual of recurrent high grade glioma patients to 
recent Mayo/NCCTG neuro-oncology trials (N997B, 987254, 967251) has been 
about 24 patients per year. In the most recent trial for recurrent GBM patients 
(N047B), 10 patients have been accrued so far between activation on 9/23/05 and 
12/01/2005. These trials have been limited to patients with only one prior 
regimen for recurrent disease, while the MV-CEA trial does not have such 
limitation. Only patients who are candidates for gross-total or subtotal tumor 
resection are eligible, however. Thus, we expect an accrual to this study of about 
8-10 patients per year, with completion of accrual in 36-42 months from study 
initiation. 
 
16.2 Analysis plans:  All the relevant results pertaining to toxicity, MTD, response, timed 
endpoints and laboratory correlates will be examined in an exploratory and hypothesis 
generating fashion. The small sample size and the heterogeneous patient population 
associated with phase I studies restricts the generalizability of the results. Any notable 
statistical result should only be viewed as an impetus for further study in phase II trials 
rather than a definitive finding in and of itself. All of the following analyses will be 
performed for Arms A and B.  
 
16.21 Adverse Events Profile 
The number and severity of all adverse events (overall, by dose-level, and by 
tumor group) will be tabulated and summarized in this patient population. The 
grade 3+ adverse events will also be described and summarized in a similar 
fashion. This will provide an indication of the level of tolerance of this treatment 
combination in this patient group. 
 
16.22 Toxicity Profile 

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As per NCI CTCAE v3.0, the term toxicity is defined as adverse events that are 
classified as either possibly, probably, or definitely related to study treatment.  
Non-hematologic toxicities will be evaluated via the ordinal CTC standard 
toxicity grading. Hematologic toxicity measures of thrombocytopenia, 
neutropenia, and leukopenia will be assessed using continuous variables as the 
outcome measures (primarily nadir) as well as categorization via CTC standard 
toxicity grading.  
 
Overall toxicity incidence as well as toxicity profiles by dose level, patient and 
tumor site will be explored and summarized. Frequency distributions, graphical 
techniques and other descriptive measures will form the basis of these analyses.  
 
16.23 Response Profile 
Best response is defined to be the best objective status 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 be determined per Section 
11.0. 
 
The number of responses is a secondary outcome measure in this trial. Responses 
will be summarized by simple descriptive summary statistics delineating 
response type (CR vs PR vs REGR), as well as stable and progressive disease in 
this patient population.   
 
Percentage of patients who are progression free at 3 and 6 months (PFS3  
and PFS6) will be similarly summarized descriptively. Time to disease 
progression and survival will be reported using standard Kaplan-Meier 
estimation method. 
 
16.24 Timed Endpoints 
The data on time-related variables will be summarized descriptively. These 
include time until any treatment related toxicity, time until treatment related 
grade 3+ toxicity, time until hematologic nadirs (WBC, ANC, platelets), time to 
progression and time to treatment failure, where time to treatment failure is 
defined as the time from registration to documentation of progression, 
unacceptable toxicity, or refusal to continue participation by the patient. 
 
16.25 Laboratory Correlates 
Descriptive statistics and simple scatterplots will form the basis of presentation 
of these data. Correlates between these laboratory values and other outcome 
measures like response and dose levels will be carried out in an exploratory 
manner. In particular, viremia, CEA titers, viral propagation in tumor, viral 
shedding, CD46 status, DTH results, CD4 and CD8 counts, LPA and ELISPOT 
results will be explored in relation to response and toxicity. 
 
16.3 
Inclusion of Women and Minorities 
 
This study will be available to all eligible patients, regardless of gender, race or ethnic 
group. There is no information currently available regarding differential agent effects in 
subjects defined by race or ethnicity. The planned analyses will, as always, look for 
differences in treatment effect based on racial groupings. The sample sizes of phase I 

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studies, however, are not sufficient to provide power for such subset analyses. 
 
To predict the characteristics of patients likely to enroll in this trial we have reviewed the 
Mayo registration classified by race. This revealed that roughly 3% of patients registered 
into cancer trials during the past five years could be classified as minorities.  
 
This would suggest that only one or two patients in the study sample are expected to be 
classified as minorities.  This precludes the possibility of a separate subset analysis 
beyond simple inspection of results for the one or two minority patients. 
 
 
17.0 
Pathology Considerations 
Central pathology review is mandatory prior to study entry to confirm eligibility. This will be 
performed by Dr. Giannini and colleagues 
.

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18.0 
Data Collection Procedure 
18.1 
Submission Time Table 
 
14 days 
after 
registrati
on 
Day 14 
after 
resectio
n 
Day 21 
after 
resectio
n4 
Day 28 
after 
resecti
on 
OB
S1 
At 
PRO
G 
Deat
h 
At Each Occurrence 
ADR 
/ 
AER 
New 
Primar
y 
Late 
Adverse 
Event 
On-Study Form 
X 
 
 
 
 
 
 
 
 
 
Baseline Adverse 
Events Form  
X 
 
 
 
 
 
 
 
 
 
Measurement 
Form 
X 
X 
X4 
X 
X1 
 
 
 
 
 
Pathology 
Reporting Form 
X 
 
 
 
 
 
 
 
 
 
Specimen 
Submission Form 
X5 
 
X4 
 
 
 
 
 
 
 
Concurrent 
Treatment Log 
X 
X 
X4 
X 
X2 
 
 
 
 
 
Nadir/Adverse 
Event Form 
 
X 
X4 
X 
X1 
 
 
 
 
 
Evaluation/Treat
ment Form 
 
X5 
X4 
 
 
 
 
 
 
 
End of Active 
Treatment Cancel 
Notification Form 
 
X 
X4 
 
 
 
 
 
 
 
Other Laboratory 
Form 
X 
 
X4 
X 
X3 
 
 
 
 
 
Evaluation/Obser
vation Form 
 
 
 
X 
X1 
 
 
 
 
 
Event Monitoring 
Form 
 
 
 
 
 
X 
X 
 
X 
X 
Secondary 
AML/MDS 
Report Form (see 
Section 10.0) 
 
 
 
 
 
 
 
X 
 
 
ADR/AER (see 
Section 10.0) 
 
 
 
 
 
 
 
X 
 
 
1. see Section 4.0 for observation schedule. 
2. Not required after 12 month post-progression evaluation. 
3. See Section 4.0 to determine if form necessary. 
4. Arm A Cohort 1 Patients only 
5. Does not apply to Arm A Cohort 1 Patients (see 4) 
 
 
19.0 
Budget Considerations 
19.1 
Costs charged to patient:  Routine clinical care. The MV-CEA will be provided free of 
charge by Mayo. However, the participant may be billed for ancillary expenses such as 
any oral medications prescribed at the time of discharge. 
 
19.2 
Tests to be research funded:  Research MRIs (or CTs), all pharmacological/correlative 

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studies, and procedures done solely for study purposes. 
 
 

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20.0 
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