PI: Fong, Timothy C Title: Novel indication for myeloid progenitor use: Induction of tolerance

Received: 12/04/2012 FOA: PA12-089 Council: 05/2013

Competition ID: ADOBE-FORMS-B2 FOA Title: PHS 2012-02 OMNIBUS SOLICITATION OF THE NIH FOR SMALL 

BUSINESS TECHNOLOGY TRANSFER GRANT APPLICATIONS (PARENT STTR 

[R41/R42])

1 R41 AI108016-01 Dual: CA,HL,NR Accession Number: 3546121

IPF: 10000154

Organization: CELLERANT THERAPEUTICS, INC.

Former Number: Department:

IRG/SRG: ZRG1 IMM-G (10)B AIDS: N Expedited: N

Subtotal Direct Costs 

(excludes consortium F&A) 

Year 1:

Animals: Y 

Humans: N 

Clinical Trial: N

Current HS Code: 10

HESC: N

New Investigator: N

Early Stage Investigator: N

Senior/Key Personnel: Organization: Role Category:

Holger Karsunky Cellerant Therapeutics PD/PI 

Adrianus Domen The Children's Mercy Hospital, Kansas 

City, MO

MPI

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State Application Identifier

Applicant Identifier

1. * TYPE OF SUBMISSION 4. a. Federal Identifier

5. APPLICANT INFORMATION * Organizational DUNS:

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If Revision, mark appropriate box(es).

9. * NAME OF FEDERAL AGENCY:

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Pre-application Application Changed/Corrected Application

Cellerant Therapeutics, inc.

1561 Industrial Road

San Carlos

94070-4111

Dr. Holger

Karsunky

Novel indication for myeloid progenitor use: Induction of tolerance

Yes No

USA: UNITED STATES

CA: California

R: Small Business

National Institutes of Health

New Resubmission

Renewal Continuation Revision

8. * TYPE OF APPLICATION:

OMB Number: 4040-0001 

Expiration Date: 06/30/2011 

b. Agency Routing Identifier

12. PROPOSED PROJECT:

* Start Date * Ending Date

06/01/2013 05/31/2014

* 13. CONGRESSIONAL DISTRICT OF APPLICANT

CA-012

14. PROJECT DIRECTOR/PRINCIPAL INVESTIGATOR CONTACT INFORMATION

* First Name: Middle Name:

* Last Name: Suffix:

Position/Title:

* Organization Name:

Department: Division:

* Street1:

Street2:

* City:

* Country: * ZIP / Postal Code:

* Phone Number: Fax Number:

* Email:

* State:

County / Parish:

Province:

Prefix:

Karsunky

Dr. Holger

94070-4111

San Carlos

1561 Industrial Road

Director, Preclinical Development

Cellerant Therapeutics

CA: California

USA: UNITED STATES

	


 		

 ! " 
SF 424 (R&R) APPLICATION FOR FEDERAL ASSISTANCE Page 2

15. ESTIMATED PROJECT FUNDING 16. * IS APPLICATION SUBJECT TO REVIEW BY STATE EXECUTIVE 

ORDER 12372 PROCESS?

a. YES a. Total Federal Funds Requested

17. By signing this application, I certify (1) to the statements contained in the list of certifications* and (2) that the statements herein are 

true, complete and accurate to the best of my knowledge. I also provide the required assurances * and agree to comply with any resulting 

terms if I accept an award. I am aware that any false, fictitious. or fraudulent statements or claims may subject me to criminal, civil, or 

administrative penalities. (U.S. Code, Title 18, Section 1001)

19. Authorized Representative

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20. Pre-application

* The list of certifications and assurances, or an Internet site where you may obtain this list, is contained in the announcement or agency specific instructions.

County / Parish:

c. Total Federal & Non-Federal Funds

18. SFLLL or other Explanatory Documentation

Province:

b. Total Non-Federal Funds

Prefix:

0.00

0.00

0.00

* I agree

Dr. Ram

Mandalam

President & CEO

Cellerant Therapeutics

1561 Industrial Road

San Carlos

CA: California

USA: UNITED STATES 94070-4111

Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

DATE:

THIS PREAPPLICATION/APPLICATION WAS MADE 

AVAILABLE TO THE STATE EXECUTIVE ORDER 12372 

PROCESS FOR REVIEW ON:

PROGRAM HAS NOT BEEN SELECTED BY STATE FOR 

REVIEW

PROGRAM IS NOT COVERED BY E.O. 12372; OR

Holger Karsunky 12/04/2012

b. NO

d. Estimated Program Income

	


 		

 ! " 
424 R&R and PHS-398 Specific

Table Of Contents Page Numbers

SF 424 R&R Face Page------------------------------------------------------------------------------------------ 1

Table of Contents--------------------------------------------------------------------------------------------- 3

Performance Sites--------------------------------------------------------------------------------------------- 4

Research & Related Other Project Information------------------------------------------------------------------ 5

 Project Summary/Abstract (Description)---------------------------------------- 6

 Public Health Relevance Statement (Narrative attachment)---------------------------------------- 7

 Facilities & Other Resources---------------------------------------- 8

 Equipment---------------------------------------- 11

Research & Related Senior/Key Person-------------------------------------------------------------------------- 12

 Biographical Sketches for each listed Senior/Key Person---------------------------------------- 13

Research & Related Budget - Year 1---------------------------------------------------------------------------- 21

 Budget Justification---------------------------------------- 24

Research & Related Budget - Cumulative Budget----------------------------------------------------------------- 26

Research & Related Budget - Consortium Budget (Subaward 1)---------------------------------------------------- 27

SBIR/STTR Information----------------------------------------------------------------------------------------- 33

PHS 398 Specific Cover Page Supplement------------------------------------------------------------------------ 35

PHS 398 Specific Research Plan-------------------------------------------------------------------------------- 37

 Specific Aims---------------------------------------- 38

 Research Strategy---------------------------------------- 39

 Vertebrate Animals---------------------------------------- 45

 Select Agent Research---------------------------------------- 48

 Multiple PI Leadership Plan---------------------------------------- 49

 Bibliography & References Cited---------------------------------------- 50

 Consortium/Contractual---------------------------------------- 55

 Resource Sharing Plan---------------------------------------- 57

PHS 398 Checklist--------------------------------------------------------------------------------------------- 58

Table of Contents Page 3

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
County:

* ZIP / Postal Code:

* Country:

* State:

* City: 

Street2:

Organization Name:

Project/Performance Site Location(s)

Project/Performance Site Primary Location

* Street1:

Province:

Cellerant Therapeutics, Inc.

1561 Industrial Road

San Carlos San Mateo

CA: California

USA: UNITED STATES

94070-4111

OMB Number: 4040-0010 

Expiration Date: 08/31/2011

I am submitting an application as an individual, and not on behalf of a company, state, 

local or tribal government, academia, or other type of organization. 

DUNS Number:

* Project/ Performance Site Congressional District: CA-012

Project/Performance Site Location a

The Children's Mercy Hospital, Kansas City, MO

2401 Gillham Road

Kansas City Jackson

MO: Missouri

USA: UNITED STATES

* ZIP / Postal Code: 64108-4619 MO-005

* Country:

Province:

* State:

* City: 

Street2:

* Street1:

Organization Name:

County:

DUNS Number:

* Project/ Performance Site Congressional District: 

I am submitting an application as an individual, and not on behalf of a company, state, 

local or tribal government, academia, or other type of organization. 

Additional Location(s) Add Attachment Delete Attachment View Attachment

	


 		

 ! " 

Performance Sites Page 4

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
1. * Are Human Subjects Involved?

IRB Approval Date:

Human Subject Assurance Number:

2. * Are Vertebrate Animals Used?

IACUC Approval Date:

Animal Welfare Assurance Number

4.b. If yes, please explain:

4.c. If this project has an actual or potential impact on the environment, has an exemption been authorized or an environmental assessment (EA) or 

 environmental impact statement (EIS) been performed?

4.d. If yes, please explain:

6. * Does this project involve activities outside of the United States or partnerships with international collaborators?

6.b. Optional Explanation:

7. * Project Summary/Abstract

11. Equipment

8. * Project Narrative

12. Other Attachments

RESEARCH & RELATED Other Project Information

Is the IACUC review Pending?

If no, is the IRB review Pending?

2.a. If YES to Vertebrate Animals

3. * Is proprietary/privileged information included in the application? 

4.a. * Does this project have an actual or potential impact on the environment? 

6.a. If yes, identify countries:

1 2 3 4 5 6

9. Bibliography & References Cited

10. Facilities & Other Resources

Yes No

1.a If YES to Human Subjects

Yes No

Yes No

If yes, check appropriate exemption number.

Yes No

Yes No

Yes No

Yes No

Yes No

Yes No

1234-ProjectSummary Final.pdf Add Attachment Delete Attachment View Attachment

1235-ProjectNarrative.pdf Add Attachment Delete Attachment View Attachment

1236-ReferencesV17HK.pdf Add Attachment Delete Attachment View Attachment

1237-Combined FacilitiesAndOtherResou Add Attachment Delete Attachment View Attachment

1238-Combined Equipment FINAL.pdf Add Attachment Delete Attachment View Attachment

Add Attachments Delete Attachments View Attachments

Is the Project Exempt from Federal regulations?

5. * Is the research performance site designated, or eligible to be designated, as a historic place? Yes No

5.a. If yes, please explain:

	


 		

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Other Information Page 5

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
PROJECT SUMMARY

State of the art techniques result in 10-year solid organ graft loss of up to 80% in cardiopulmonary organ 

transplantation, and re-transplantation is often not possible. Establishment of donor-specific immunological 

tolerance (DSIT), a condition in which a recipient accepts a transplant without immunosuppression, while 

retaining the ability to fight infections, would reduce graft loss. The only identified method of inducing robust 

tolerance involves Hematopoietic Cell Transplantation (HCT), usually in the form of bone marrow 

transplantation (BMT). Though long recognized experimentally as a means of inducing DSIT, clinical 

translation has been limited due to the associated complications. A major problem has been the treatment 

necessary to prepare a recipient for a blood cell transplant. White blood cell counts fall precipitously, resulting 

in neutropenia and increased susceptibility to infections. To reduce infections associated with neutropenia in 

HCT recipients, unrelated myeloid progenitors (MP) can be injected together with the HCT. This therapy is 

effective in the laboratory setting in reducing deaths caused by bacterial and fungal infections, and several 

trials testing a clinical MP product developed by Cellerant Therapeutics (CLT-008) MP in humans are ongoing. 

We have discovered that injection of MP under these conditions results in MP-specific tolerance, even though 

there may be only very low-level MP engraftment after the first month. Important, MP from B10;B6-Rag2-/-Il2rg-

/- mice, which are incapable of producing functional B, T or NK cells, induce tolerance and clearly show that 

organ graft-matched lymphoid cells are not essential under these conditions. Uniquely, MP cells induce 

antigen-specific tolerance in our experimental model system, are clinically available, and have been 

associated with minimal to no side effects in current clinical trials. MP constitute an ideal and 

innovative approach in tolerance induction protocols, preferred over efforts aimed at achieving high 

level donor chimerism.

 The proposed research in phase I will focus on (Aim 1) testing whether MP can prevent specific lung 

transplant rejection symptoms, important because lung transplants may be a good patient population for initial 

trials and (Aim 2) on testing the degree of mismatch allowed between MP and organ graft, as this will affect the 

production of clinical grade MP for clinical tolerance trials. The design of these trials using a Cellerant 

produced clinical MP product would be the next stage following this STTR project. 

Project Description Page 6

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
PROJECT NARRATIVE 

The proposal aims to develop methods that can be used to improve the long-term outcome of clinical organ 

transplantations. State of the art technology results in up to 80% organ graft loss over ten years mostly due to 

rejection, and re-transplantation is often not possible. The academic partner has developed preclinical 

tolerance models and has shown that myeloid cells can induce specific tolerance. The small business partner 

is currently studying an innovative product (CTL-008, human Myeloid Progenitor Cells) in clinical trials for the 

prevention of infections. Together we are aiming to test if a commercial Myeloid Progenitor Cell product for the 

induction of tolerance in solid organ transplantation is feasible and to transfer this technology to the small 

business partner for further clinical development and application such that it can improve the long-term 

outcome for patients undergoing transplantation for end-stage organ failure. 

Public Health Relevance Statement Page 7

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
FACILITIES AND OTHER RESOURCES

1) At Cellerant Therapeutics (Laboratory of Dr. Karsunky)

Facilities: Cellerant has ample of space and adequate facilities to accommodate the proposed work. Cellerant 

has approximately 42,000 sq. ft. total of laboratory, animal vivarium, office, and common area space in San 

Carlos, CA. The facility consists of twenty two laboratories and a vivarium.

Laboratory: Ten laboratories are dedicated tissue culture suites for mouse, human or NHP tissue culture. 

Each of them is equipped with 1-3 biological safety cabinets, 2-6 incubators, centrifuge, microscope and other 

tissue culture related equipment. All rooms are supplied with HEPA filtered air. Seven laboratories are 

multipurpose labs with multiple wet benches for molecular biology, PCR, protein work, histology, 

immunochemistry, QC and other techniques. Two laboratories are dedicated to process and assay 

development work for cell culture processes and contain bioreactors, gas and metabolic analyzers, etc. One 

laboratory contains equipment for flow cytometry and cell sorting. This laboratory also contains three biological 

safety cabinets and multiple incubators dedicated to non-human tissue culture and processing. Two are 

dedicated microscope rooms. 

In addition, there are four freezer rooms that house the liquid nitrogen cryogenic freezers, -150°c ultra 

low mechanical freezers and controlled rate freezers for cell cryopreservation and stability studies. One 

dedicated glass wash room with autoclave and glassware washer and one 4°c walk-in cold room. Incubators, 

refrigerator and freezers are monitored. Critical equipment undergoes regularly scheduled preventive 

maintenance and calibration or certification. The facilities are back up with emergency generators.

Animal: Cellerant has approximately 3,700 sq. ft. animal facility with a separate entry from the laboratories for 

the housing and maintenance of mouse colonies. The vivarium consists of four housing rooms, two procedure 

rooms, one quarantine room, a dirty and clean wash/preparation room, including an autoclave, cage rack 

washer, bottle filler and an X-ray irradiation room for whole body irradiation of mice. The facility includes a 

changing room, protective gowning room, toilet and shower. This facility can house approximately 5,000 mice 

and its environment is controlled and monitored for temperature, humidity and light cycles. Entry into the 

building and vivarium is controlled by electronic key card. 

Office: The remaining space houses offices, cubicles and workstations, conference, computer server room, 

electronic key card documentation room and break rooms. All staff have their own computer work station that 

are connected network. All computers are equipped with MS Office and other work relevant software. 

2) At Children’s Mercy Hospital/University of Missouri, Kansas City (Laboratory of Dr. Domen)

Facilities: Part of the proposed research will be performed at Children’s Mercy Hospital and Clinics in Kansas 

City where the lab has 600 sq ft of wet lab space and 100sq ft of office space for the PI. In addition there is 

access to the animal facilities at UMKC to allow with shared access to a mouse room as well as two procedure 

rooms. The animal space is located in the school of Pharmacy across the road from Children’s Mercy 

Hospital. 

The Ward Family Center for Congenital Heart Disease has a uniquely developed research and 

informatics section. The academic and research section has six full time employees and one part time 

employee dedicated strictly to the administration of research projects. This group currently administers 25 

cardiac IRB protocols, as well as 6 IACUC protocols, and 12 IBC protocols. Two of the cardiovascular 

operating room suites within Children's Mercy Hospitals and Clinics are equipped with a GMP grade clean 

room connected directly between the suites. This state-of-the-art facility was specifically designed to handle 

bio-engineered tissue, and allow for the implantation of this tissue in the clinical setting. 

Laboratory: The 600sq ft research laboratory is equipped with the basic equipment found in modern 

biomedical research laboratories including microscopes, refrigerated centrifuges, PCR machines, and -20 and 

Facilities Page 8

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
-80 freezers. In addition a 8 parameter Veterinary blood cell counter (ScilVetABC) is available in the lab Full 

capabilities for tissue culture exist. Major equipment is shared among the laboratories. Major equipment 

includes a CMH-owned RadSource 2000 irradiator in the mouse facility at the UMKC medical school where the 

mouse facilities used for this project are located and a 6 color Blue/Red laser Attune Flow Cytometer located in 

the Domen lab. 

Clinical: The clinical laboratory is a full service laboratory operated by the Department of Pathology which 

occupies 8,000 ft2

 on the second floor of the hospital and provides laboratory services to the hospital and 

ambulatory clinics 24 hours/day, 7 days per week. Areas of clinical service provided include routine and special 

chemistry, endocrinology, flow cytometry, coagulation, hematology, histology, diagnostic immunology, 

microbiology, transfusion services, toxicology/TDM, in vitro nuclear medicine, cytogenetics, biochemical 

genetics and urinalysis. The laboratory is fully accredited by CAP #19365-01 and licensed by CLIA 

#26DO443323. It is capable of providing clinical laboratory testing required for clinical research protocols.

Animal: Animal facilities are available through a cooperative agreement with the University of MissouriKansas City. Facilities (AAALAC certified) are provided for small and large animals with the animal facility 

directed by a veterinarian. Equipment essential to the proposed experiments, most notably a RadSource 2000 

mouse x-ray irradiator, are present. The animal facility is located across the road from Children’s Mercy.

Computer: All hospital staff are connected to a Children’s Mercy Hospital client/server LAN with word 

processing, spreadsheet, database, electronic mail, presentation, and graphical software. In addition, 

Research and Grants Administration has a color scanner with OCR and graphics software, a HP 1000C color 

printer and laser printers. A full range of scientific graphical (Sigma Plot v 4.0) and statistical packages (SAS, 

SPSS, S+, StatXact, and DBMS-Copy) are available. On-line library/literature search software, including 

Knowledge Server for NLM access, End Note bibliographic software, and Current Contents is available on the 

network. A fractional DS-3 line provides high speed, firewall-protected access to the Internet. 

Users logging onto the system have access to a personal directory on a server that is backed up daily. The 

network is maintained by the Hospital Information Services.

A public site as well as an internal Research and Grants Tools Portal website with links to internal CMH 

and external websites (http://www.childrensmercy.org/MedicalResearch.aspx) is maintained to provide 

assistance to researchers. 

Members of the lab are equipped with iMac computers running system 10.6 or 10.5.

Office: A 100 sq ft office (3730.08) equipped with high speed internet, a 27inch iMac computer (System 10.6) 

with appropriate software (Microsoft Office, iWorks, Canvas, Filemaker, Endnote, Adobe Acrobat, Graphpad 

Prism, OmniGraffle Pro) and a color laser printer and scanner is available for the PI, located next to the wet lab 

space. 

Interactions: Members of the laboratory participate in the regular meetings and interactions of the larger 

Immunology community that brings together researchers at the University of Kansas Medical Center, the 

University of Kansas Lawrence Campus and Children’s Mercy Hospital 

(http://www.kumc.edu/immunology.html), bringing together researchers addressing a wide range of 

immunological questions.

Other: Library: The CMH library is located on the ground floor of the hospital. Three hundred and twenty-five 

current periodical subscriptions and 3,900 bound volumes are maintained. Library staff provides online access 

to the NLM (MEDLINE) and BRS. Interlibrary loan is readily available. Adjacent to the hospital, the University 

of Missouri-Kansas City (UMKC) School of Medicine Library maintains 722 periodical subscriptions, 71,666 

bound volumes and faculty support services.

The UMKC library system contains 1,005,724 volumes, 8,868 current serial subscriptions, 1,876,258 

microfilms, and 797,602 government documents. The libraries have major access agreements with CRL, NLM, 

UM OVID data bases, UM SiteSearch databases, OCLC FirstSearch Basic, RLIN/BIB Eureka, ICPSR, and 

MIRACL.

Facilities Page 9

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
UMKC medical and health sciences resources are supplemented by the nearby libraries of the University of 

Kansas Medical Center Archie Dykes Library.

Biomedical: Children's Mercy Hospital provides a machine, electrical and biomedical shop to service technical 

equipment.

Facilities Page 10

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
EQUIPMENT

1) At Cellerant Therapeutics (Laboratory of Dr. Karsunky)

Cellerant has already on-site all major laboratory equipment needed to complete the proposed work. 

For tissue culture work a total of 19 biosafety cabinets and 30 humidified CO2 incubators are currently installed 

and operational. Two of the incubators include triple gas controls for low oxygen conditions. The flow cytometry 

lab is equipped with two Becton Dickinson FACSAria II flow cytometer cell sorters, one Becton Dickinson 

FACSCalibur flow cytometer, one Beckman Coulter Gallios and one Dako Cyan flow cytometer. For magnetic 

cell isolation one Miltenyi AutoMACS and two AutoMACSpro as well as two Isolex 300i cell isolators and a 

Miltenyi CliniMACS cell isolator are at disposal. For storage the labs are equipped with multiple refrigerators 

and freezers for reagent storage (2-8°, -20° and -80°C), six liquid nitrogen tanks for cryogenic cell storage, and 

one control rate freezer. Other equipment includes one Coulter hematology analyzer for human blood, a 

Hemavet 950S hematology analyzer for animal blood, one microtome for sectioning, two multi-well plate 

readers with automated plate washer, two UV Vis spectrophotometers, one NucleoCounter and one Vi-Cell cell 

counter, and an Alpha Innotech ChemiImager. The process development lab houses a Wave bioreactor, 

Micro24 mini bioreactor system, a Haemonetics PCS2 cell collection system, an YSI bionalyzer and a 

StatProfile pHOX gas analyzer. Additional bench top laboratory equipment includes several bench top 

centrifuges with rotors, microcentrifuges, PCR machines, various microscopes, water baths and several 

electrophoresis units. 

The vivarium contains numerous cage racks with single ventilated cages, one pass-through cage 

washer, one large autoclave, one water bottle washer, one water bottle filler. Every holding room is equipped 

with a cage changing station. One procedure room is equipped with a biosafety cabinet for surgical 

procedures including an isoflurane anesthesia machine. The radiation chamber houses one Faxitron CP-160 

X-ray irradiator with an output of 72 cGy/min that can irradiate up to ten mice at a time. 

Cellerant performs routine preventive maintenance and regular calibration on its critical equipment. 

All occupied offices and cubicles are equipped with personal computers linked to a local network. 

Electronic data are archived daily and copies stored off-site.

2) At Children’s Mercy Hospital/University of Missouri, Kansas City (Laboratory of Dr. Domen)

Research laboratories are individually equipped with the basic equipment found in modern biomedical research 

laboratories including microscopes, refrigerated centrifuges, PCR machines, and very low temperature 

freezers. In addition a 8 parameter Veterinary blood cell counter (ScilVetABC) is available in the Domen lab. 

Full capabilities for tissue culture exist. 

Major equipment is shared among the laboratories. Major equipment includes a CMH-owned 

RadSource 2000 irradiator in the mouse facility at the UMKC medical school where the mouse facilities used 

for this project are located.

Major equipment at CMH includes a Applied Biosciences 6-color Blue/Red laser Attune Flow Cytometer 

with plate reader located in the Domen Lab, Beckman UV-Visible spectrophotometer, Applied Biosystems 

Model 3100 Capillary electrophoresis-based separation and identification system, Agilent Bioanalyzer 2100, 

Molecular Dynamics STORM 860 imaging system, ABI Prism 7000 quantitative real-time PCR equipment, 

Beckman computer-controlled spectrophotometer, Savant Speed Vac vacuum centrifuge, Applied Biosystems 

automated nucleic acid purifier, and Amersham two laser imaging system for fluorescence and 

phosphorescence. A Paradigm ultrasound backscatter microscope is available to directly visualize in utero

animal embryos that permits the introduction of agents to alter the course of organ formation.

Equipment Page 11

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Province:

PROFILE - Project Director/Principal Investigator

Prefix: * First Name: Middle Name:

* Last Name: Suffix:

Organization Name: Division:

Position/Title: Department:

* Street1:

Street2:

* Phone Number: Fax Number:

* E-Mail:

Credential, e.g., agency login:

* Project Role: Other Project Role Category:

* Country: * Zip / Postal Code:

* State:

* City: County/ Parish:

Attach Current & Pending Support

RESEARCH & RELATED Senior/Key Person Profile (Expanded)

*Attach Biographical Sketch

Dr. Holger

Karsunky

Director, Preclinical Development

Cellerant Therapeutics

1561 Industrial Road

San Carlos

CA: California

USA: UNITED STATES 94070-4111

PD/PI

PhD

2000

1244-biosketch Karsunky 2012 Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

OMB Number: 4040-0001 

Expiration Date: 06/30/2011

Degree Type:

Degree Year:

Province:

PROFILE - Senior/Key Person

Prefix: * First Name: Middle Name:

* Last Name: Suffix:

Organization Name: Division:

Position/Title: Department:

* Street1:

Street2:

* Phone Number: Fax Number:

* E-Mail:

Credential, e.g., agency login:

* Country: * Zip / Postal Code:

* State:

* City: County/ Parish:

Prof. G.W.

Assistant Professor

The Children's Mercy Hospital, Kansas City, MO

Jackson

Pediatrics

USA: UNITED STATES

Adrianus

Domen

2401 Gillham Road

Kansas City

MO: Missouri

64108-4619

* Project Role: PD/PI Other Project Role Category:

1

Degree Type: PhD

Degree Year: 1993

*Attach Biographical Sketch 1245-BiosketchJD_2012DomenSTT Add Attachment Delete Attachment View Attachment

Attach Current & Pending Support Add Attachment Delete Attachment View Attachment

	


 		

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Key Personnel Page 12

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
BIOGRAPHICAL SKETCH 

Provide the following information for the key personnel and other significant contributors in the order listed on Form Page 2. 

Follow this format for each person. DO NOT EXCEED FOUR PAGES.

NAME 

Holger Karsunky

eRA COMMONS USER NAME 

POSITION TITLE 

Director, Preclinical Development 

EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, and include postdoctoral training.)

INSTITUTION AND LOCATION DEGREE 

(if applicable) YEAR(s) FIELD OF STUDY 

University of Marburg, Germany M.Sc. 1991-1996 Molecular Biology 

University of Essen Medical School, Germany Ph.D. 1996-2000 Immunology 

University of Essen Medical School, Germany Postdoctoral 

Fellow 2000-2001 Hematopoiesis and T 

cell development 

Stanford University, California Postdoctoral 

Fellow 2001-2005 Hematopoietic stem 

cells and progenitors 

A. Personal Statement 

For the last 10+ years I have been working on the biology of myeloid progenitor cells and their potential clinical 

application. Since joining Cellerant the focus of my work has been the development of CLT-008, human 

Myeloid Progenitor Cells, which is in advanced clinical development. The use of myeloid progenitor cells offers 

the opportunity to have a novel off-the-shelf cell therapeutic that can be stored long term and used without 

recipient HLA matching. CLT-008 is currently developed for three indications: 1) to enhance hematopoietic 

recovery in patients receiving umbilical cord blood transplantation after myeloablative conditioning; 2) to 

prevent infection in leukemia patients receiving high dose chemotherapy; and 3) as a medical countermeasure 

for acute radiation syndrome. Through the combination of safety studies in animal xenograft models and proofof-concept efficacy studies using a comparable animal product my group was able to demonstrate its safety 

and efficacy in animal models. Cellerant subsequently filed an investigational new drug (IND) application and 

has since then initiated two currently ongoing clinical trials. I have extensive experience in the field of human 

and mouse hematopoietic stem and progenitor cell biology. As a postdoctoral fellow at Stanford I worked on 

the characterization, lineage relationship and developmental potential of myeloid and lymphoid progenitor 

populations. I am very familiar with the regulatory requirements for cell based therapies and have led as the 

responsible PI or Program Director similar and significantly larger research projects. In response to Program 

Announcement PA-12-089 together with my colleagues Dr. Adrianus (Jos) Domen we are proposing to 

conduct proof-of-concept preclinical work to test the hypothesis if Cellerant’s Myeloid Progenitor technology 

can be used to induce tolerance in patients receiving allogeneic solid organ transplants. Based on the initial 

work performed in Dr. Domen’s lab I believe it is feasible to develop a modified version of CLT-008 specific for 

the induction of tolerance which could have a significant impact on the clinical outcome of many transplants. 

B. Positions and Honors 

Positions and Employment

2011-current Director, Preclinical Development, Cellerant Therapeutics, San Carlos, CA 

2009-2011 Associate Director, Development, Cellerant Therapeutics, San Carlos, CA 

2008-2009 Group Leader, Development, Cellerant Therapeutics, San Carlos, CA 

2007-2008 Senior Staff Scientist, Cellerant Therapeutics, San Carlos, CA 

2005-2006 Research Staff Scientist, Cellerant Therapeutics, San Carlos, CA 

2001-2005 Post-doctoral fellow, Stanford University School of Medicine, Department of Pathology (PI, Dr. 

Irving L. Weissman) 

2000-2001 Post-doctoral fellow, University of Essen School of Medicine, Institute for Cell Biology and 

Cancer Research (PI, Dr. Tarik Möröy) 

1996-2000 Graduate student, University of Essen School of Medicine, Institute for Cell Biology and 

Cancer Research (PI, Dr. Tarik Möröy) 

1995-1996 Graduate student, University of Marburg (Dr. Tarik Möröy) 

Biosketches Page 13

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Other Experience and Professional Memberships 

2009- Member International Society of Stem Cell Research 

2010- Member Radiation Research Society 

Honors 

2001 – Postdoctoral Fellowship of the Ernst Schering Research Foundation 

2000 – Earned doctoral degree with summa cum laude from the University of Essen 

1996 – Doctoral Fellowship of the German Research Foundation (DFG) 

C. Selected Peer-reviewed Publications (Selected from 37 peer-reviewed publications)

1. Singh, VK, Christensen J, Fatanmi OO, Gille D, Ducey EJ, Wise SY, Karsunky H, and Sedello AK. 

Myeloid progenitors: a radiation countermeasure that is effective when initiated days after irradiation 

Radiat Res, 177: 781-791, 2012. 

2. Fathman. J, Bhattacharya D, Inlay M, Seita J, Karsunky H, and Weissman IL. Identification of the 

earliest transplantable Natural Killer cell committed progenitor in murine bone marrow. Blood 118: 5439-

5447, 2011. 

3. Ji H, Ehrlich L, Seita J, Murakami P, Doi A, Lindau P, Lee H, Aryee M, Irizarry R, Kim K, Rossi D, Inlay M, 

Serwold T, Karsunky H, Ho L, Daley G, Weissman IL and Feinberg, AP. A comprehensive methylome 

map of lineage commitment from hematopoietic progenitors. Nature 467: 338-342, 2010. 

4. Papathanasiou P, Attema JL, Karsunky H, Hosen N, Sontani Y, Hoyne GF, Smale ST, and Weissman 

IL. Self-renewal of the long-term reconstituting subset of hematopoietic stem cells is regulated by Ikaros. 

Stem Cells 27, 3082-3092, 2009. 

5. Inlay MA, Bhattacharya D, Sahoo D, Serwold T, Seita J, Karsunky H, Plevritis SK, Dill DL, and 

Weissman IL. Ly6d marks the earliest stage of B cell specification and identifies the branchpoint between 

B cell and T cell development. Genes & Development 23: 2376-81, 2009. 

6. Papathanasiou P, Attema JL, Karsunky H, Xu J, Smale ST, and Weissman IL. Evaluation of the LongTerm Reconstituting Subset of Hematopoietic Stem Cells with CD150. Stem Cells 27: 2498-2508, 2009. 

7. Ooi AG, Karsunky H, Majeti R, Butz S, Vestweber D, Ishida T, Quertermous T, Weissman IL, and 

Forsberg EC. The Adhesion Molecule ESAM1 is A Novel Hematopoietic Stem Cell Marker. Stem Cells 

27: 653-661, 2008. 

8. Cao Y, Wagers AJ, Karsunky H, Zhao H, Reeves R, Wong RJ, Stevenson DK, Weissman IL, and 

Contag CH. Heme Oxygenase 1 Deficiency Compromises Stress Responses of Hematopoietic Stem 

Cells. Blood 112: 4494-4502, 2008. 

9. Karsunky H, Inlay, MA, Serwold TF, Bhattacharya D, and Weissman IL. Flk2+

 common lymphoid 

progenitors possess equivalent differentiation potential for the B and T lineages. Blood 111: 5562-5570, 

2008. 

10. Frontelo MP, Manwani D, Galdass M, Karsunky H, Gallagher PG, and Bieker JJ. Novel role for EKLF in 

megakaryocyte-erythroid lineage differential commitment. Blood 110: 3871-3880, 2007. 

11. Fernandez I, Zeiser R, Karsunky H, Kambham N, Soderstrom K, Negrin RS and Engleman EG. CD101 

surface expression discriminates potency among murine FoxP3+

 regulatory T cells. J. Immunol. 179: 

2808-2814, 2007. 

12. Sanyal M, Tung JW, Karsunky H, Zeng H, Selleri L, Weissman IL, Herzenberg LA, and Cleary ML. B cell 

development fails in the absence of the Pbx1 proto-oncogene. Blood 109: 4191-4199, 2007. 

13. Mende I, Karsunky H, Weissman IL, Engleman EG, and Merad M. Flk2+

 myeloid progenitors are the 

main source of Langerhans cells during inflammatory conditions. Blood 107: 1383-1390, 2006. 

Biosketches Page 14

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
14. Karsunky H, Merad M, Mende I, Manz MG, Engleman EG, and Weissman IL. Ontogeny of Interferon 

alpha Producing Dendritic Cells. Exp. Hematol. 33: 173-181, 2005. 

15. So CW, Karsunky H, Wong P, Weissman IL, and Cleary M. Leukemic transformation of hematopoietic 

progenitors by MLL-GAS7 in the absence of Hoxa7 or Hoxa9. Blood 103: 3192-3199, 2004. 

16. Cozzio A, Passegue E, Ayton PM, Karsunky H, Cleary ML, and Weissman IL. Similar MLL-associated 

leukemias arising from self-renewing stem cells and short-lived myeloid progenitors. Genes & 

Develop.17: 3029-3035, 2003. 

17. Adhikary S, Peukert K, Karsunky H, Lutz W, Elsässer HP, Möröy T, and Eilers M. Miz1 is required for 

early embryonic development during gastrulation. Mol. Cell Biol. 23: 7648-7657, 2003 

18. Karsunky H, Merad M, Cozzio A, Weissman IL, and Manz MG. Flt3 ligand regulates dendritic cell 

development from Flt3-positive lymphoid and myeloid committed progenitors to Flt3-positive dendritic 

cells in vivo. J. Exp. Med., 198: 305-313, 2003. 

19. Yücel R, Karsunky H, Klein-Hitpass L, and Tarik Möröy. The transcriptional repressor Gfi1 affects 

development of early, uncommitted c-Kit+ T-cell progenitors and CD4/CD8 lineage decision in the 

thymus. J. Exp. Med., 197: 831-844, 2003. 

20. So CW, Karsunky H, Passegue E, Cozzio A, Weissman IL, and Cleary ML. MLL-GAS7 transforms 

multipotent hematopoietic progenitors and induces mixed lineage leukemias in mice. Cancer Cell, 3: 161- 

71, 2003. 

21. Geisen C, Karsunky H, Yücel R, and Möröy T. T-cell lymphoma in CD2-cyclin E transgenic mice that are 

deficient for p27Kip1. Oncogene 22: 1724-1729, 2003. 

22. Merad M, Manz MG, Karsunky H, Wagers A, Peters W, Charo I, Weissman IL, Cyster JG, and 

Engleman EG. Langerhans cells renew in the skin throughout life under steady-state conditions. Nat. 

Immunol. 3:1135-41, 2002 

23. Karsunky H , Zeng H, Schmidt T, Zevnik B, Kluge R, Schmid KW, Dührsen U, and Möröy T. 

Inflammatory reactions and severe neutropenia in mice lacking the transcriptional repressor Gfi1. Nature 

Genetics 30: 295-300, 2002. 

24. Karsunky H, Mende I, Schmidt T, and Möröy T. High levels of the onco-protein Gfi-1 accelerate T-cell 

proliferation and inhibit activation induced T-cell death in Jurkat T-cells. Oncogene 21: 1571-1579, 2002. 

25. Staller P, Peukert K, Kiermaier A, Seoane J, Lukas J, Karsunky H, Möröy T, Bartek J, Massague J, 

Häne F, and Eilers M, Repression of p15INK4b expression by Myc through association with Miz-1. 

Nature Cell Biol. 3: 392-399, 2001. 

26. Rödel B, Tavassoli K, Karsunky H, Schmidt T, Schaper F, Heinrich P, Shuai K, Elsässer HP, and Möröy 

T. The zinc finger protein Gfi-1 can enhance STAT3 signaling by interacting with the STAT3 inhibitor 

PIAS3. EMBO J. 19: 5845-5855, 2000. 

27. Beier R, Burgin A, Kiermaier A, Fero M, Karsunky H, Saffrich R, Möröy T, Ansorge W, Roberts J, and 

Eilers M. Induction of cyclin E-cdk2 kinase activity, E2F-dependent transcription and cell growth by Myc 

are genetically separable events. EMBO J. 19: 5813-5823, 2000. 

28. Napirei M, Karsunky H, Zevnik B, Stephan H, Mannherz HG, and Möröy T. Systemic Lupus 

Erythematosus (SLE) in Dnase 1 deficient mice. Nature Genetics 25: 177-181, 2000. 

29. Karsunky H, Geisen C, Schmidt T, Zevnik B, Gau E, and Möröy T. Oncogenic potential of cyclin E in 

Tcell lymphomagenesis in transgenic mice: Evidence for cooperation between cyclin E and Ras but not 

Myc. Oncogene 18: 7816-7825, 1999. 

30. Schmidt T, Karsunky H, Zevnik B, Elsässer HP, and Möröy T. Zinc fingerprotein Gfi-1 has low oncogenic 

potential but cooperates strongly with Pim and Myc genes in T-cell lymphomagenesis. Oncogene 17: 

2661-2668, 1998. 

Biosketches Page 15

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
31. Schmidt T, Karsunky H, Rödel B, Zevnik B, Elsässer HP, and Möröy T. Evidence implicating Gfi-1 and 

Pim-1 in pre T-cell differentiation steps associated with beta-selection. EMBO J. 17: 5349-5359, 1998. 

32. Haas K, Johannes C, Geisen C, Schmidt T, Karsunky H, Blass-Kampmann S, Obe G, and Möröy T. 

Malignant transformation by cyclin E and Ha-ras correlates with resistance against cell death but requires 

functional Myc and CDK4. Oncogene 15: 2615-2624 1997. 

33. Zörnig M, Schmidt T, Karsunky H, Grzeschiczek A, and Möröy T. Zinc finger protein GFI-1 cooperates 

with myc and pim-1 in T-cell lymphomagenesis by reducing the requirements for IL-2. Oncogene 12: 

1789-801, 1996. 

D. Research Support. 

Ongoing Research Support

HHSN261200100076C Karsunky (PI) 09/20/12 – 06/19/13 

Development of a Process for Generating Human MKP. 

Role: PI 

Time effort: 5% 

HHSO100201000051C Mandalam (PI) 09/01/10 – 08/31/13 

Advanced Therapeutics for Treating Neutropenia Resulting from Acute Exposure to Ionizing Radiation 

Role: Program Director 

Time effort 90% 

Completed Research Support

1 RC1AI080314-01S1 Karsunky (PI) 09/17/09 - 08/31/11 

Development of ex vivo Expanded Megakaryocyte Progenitors for Platelet Recovery 

Role: PI 

Time effort: 10% 

HHSO 100200800063C Mandalam (PI) 09/15/08 – 08/31/11 

Development of human Myeloid Progenitor Cells, CLT-008, as a countermeasure for the treatment of Acute 

Radiation Induced Hematopoietic Syndrome 

Role: Leading Investigator 

Time effort 40% 

1 RC1AI080314-01 Karsunky (PI) 07/01/08 - 12/31/09 

Development of ex vivo Expanded Megakaryocyte Progenitors for Platelet Recovery 

Role: PI 

Time effort: 50% 

1 R43 AI064156-01 Karsunky (PI) 06/15/05 - 06/14/06 

Application of Expanded Progenitors against Infection 

Role: PI 

1 R43 AI061856-01 Karsunky (PI) 07/01/2004 – 12/31/2006 

Expansion of HSC for rescue in biodefense applications 

Role: PI 

Biosketches Page 16

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
BIOSKETCHES for the Principal Investigator and all Co-Investigators 

BIOGRAPHICAL SKETCH 

Provide the following information for the Senior/key personnel and other significant contributors. Follow this format for each person. DO NOT 

EXCEED FOUR PAGES. 

NAME 

Adrianus (Jos) G.W. Domen 

eRA COMMONS USER NAME (credential, e.g., agency login) 

POSITION TITLE 

Lab manager, Cardiac Transplant Lab, CMH 

Assistant Professor, University of Missouri Kansas 

City 

EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and 

residency training if applicable.) 

INSTITUTION AND LOCATION DEGREE 

(if applicable) 

MM/YY FIELD OF STUDY 

University of Nijmegen M.Sc. 1978-1985 Biology 

Netherlands Cancer Institute, University of 

Amsterdam Ph.D. 1985-1993 Oncology 

Stanford University Postdoctoral 1993-1999 Immunology/Stem Cell 

Biology 

A. Personal Statement 

The aim of the proposed research is to use myeloid progenitors as a cellular therapeutic for tolerance 

induction. The proposed research describes preclinical studies necessary to device an approach that can be 

tested in the clinical realm, and I have the experience and capabilities to bring this project to a successful 

completion. My background in research using mouse models is extensive. I obtained expertise in mouse 

hematopoietic system studies in multiple environments including the Netherlands, the UK (Manchester) and 

the USA (Stanford, Duke and in Biotechnology). This experience dates as far back as my thesis work at the 

Netherlands Cancer Institute during which I studied lymphomagenesis and performed analysis of oncogenenull mutant mice and transgenic mice. I extended this as a postdoctoral fellow at Stanford and faculty member 

at Duke to more basic Hematopoietic Stem Cell biology. While at Cellerant Therapeutics, I began focusing on 

Myeloid Progenitor cells in work that was supported by an NIH SBIR grant, specifically on obtaining clinically 

useful amounts of cells using short term cultures of mouse and human hematopoietic stem cells and testing 

efficacy of the expanded in mouse infection models. The program that I initiated has since garnered major 

government support and has progressed to successful clinical trials. In a longstanding collaboration with my 

wife, Kimberly Gandy, many of our lab efforts have become focused on the use of HCT to induce tolerance. 

Tolerance is a longstanding area of interest to us, uniquely suiting her dual background (Cardiac and lung 

transplant surgeon with a PhD in immunology/stem cell biology). 

 Due to the training/job requirements of my wife the lab has been relocated four times in a period of a 

little over six years (2004-2010), significantly reducing productivity. However, during the three years in 

Milwaukee, supported by outside funding (including a 2 year grant from Advancing Healthier Wisconsin), the 

foundation was laid for the current work in Kansas City. The myeloid progenitor/tolerance induction work has 

been published in 2011. In addition, one manuscript is currently submitted and another is in the final stages of 

preparation. We are confident that in direct collaboration with Cellerant Therapeutics we can help accelerate 

this work to the clinical test stage, necessary for translation to the clinic. 

B. Positions and Honors 

Positions and Employment

2000–2004 Assistant Research Professor of Medicine, Dept. of Medicine, Duke University, Durham NC 

2001–2004 Assistant Research Professor of Immunology, Dept. of Immunology, Duke University 

2004–2005 Staff Scientist, Cellerant Therapeutics Inc., Palo Alto, CA 

2005–2007 Research Assistant Professor of Surgery, Dept. of Surgery, University of Arizona, Tucson 

2007–2010 Assistant Professor of Surgery, Dept. of Surgery, Medical College of Wisconsin, Milwaukee 

2008–2010 Assistant Professor of Surgery and Cell Biology, Neurobiology & Anatomy, MCW 

2011-present Lab manager, Cardiac Transplant Research Lab, Children’s Mercy Hospital 

2011-present Assistant Professor, Dept of Pediatrics, University of Missouri Kansas City 

Biographical Sketches for each listed Senior/Key Person 2 Page 17

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Other Experience and Professional Memberships

1995–present American Association for the Advancement of Science 

1998–present American Society of Hematology 

1998–present American Association of Immunologists 

2000–present International Society for Stem Cell Research 

2003–present American Society for Bone Marrow Transplantation 

Patents: 

Methods and compositions for modulating lifespan of hematolymphoid cells. 

 Weissman, I.L., Lagasse, E. and Domen, A.G.W. US Patent 5,614,397 

Methods of expanding Myeloid Cell Populations and uses thereof. 

 Fong, T., Domen, A.G.W. and Christensen, J.L. US Patent App: 20060134783. 

Honors

1989 Short-Term EMBO Fellowship 

1993–1994 Dutch Cancer Foundation Fellowship 

1992 Invited lecture: Modern Trends in Human leukemia, Wilsede X. Leukemia 7:s108-s112(1993) 

2001 Kimmel Scholar Award 

C. Selected Peer-reviewed Publications (Selected from 44 published) 

Most relevant to the current application

• Domen, J., Sun, L., Trapp, K., Maghami, N., Inagaki, E., Li, Y., Simpson, P. and Gandy, K.L. Tolerance 

induction by hematopoietic cell transplantation: combined use of progenitor and stem cells. J. Heart Lung 

Transplant, 30:507-514, 2011. PMID: 21256050 

• Domen, J., Gandy, K.L. and Dalal, J. Emerging uses for pediatric hematopoietic stem cells. Pediatric Res 

71:411-417 (2012). PMID: 22278186

• Domen J., Gandy K.L., Weissman I.L. Systemic overexpression of BCL-2 in the hematopoietic system 

protects transgenic mice from the consequences of lethal irradiation. Blood 91:2272–2282, 1998. PMID: 

9516125. 

• Gandy K.L., Domen, J., Aguila L. and Weissman I.L. CD8+

TCR+ and CD8+TCR- cells in whole bone 

marrow facilitate the engraftment of HSC across allogeneic barriers. Immunity 11:579–590, 1999. PMID: 

10591183. 

• Domen J., Cheshier S.H., and Weissman, I.L. The role of apoptosis in the regulation of hematopoietic 

stem cells; overexpression of BCL-2 increases both their number and repopulation potential. J Exp Med

191:253–263, 2000. PCMID: PMC2195763. 

Additional recent publications of importance to the field (in chronological order)

• Saris C.J., Domen J., Berns A. The pim-1 oncogene encodes two related protein-serine/threonine kinases 

by alternative initiation at AUG and CUG. Embo J 10:655–664, 1991. PMCID: PMC452698. 

• Domen J., van der Lugt N.M., Laird P.W., Saris C.J., Clarke A.R., Hooper M.L., Berns A. Impaired 

interleukin-3 response in Pim-1-deficient bone marrow-derived mast cells. Blood 82:1445–1452, 1993. 

PMID: 7689870. 

• Domen J., van der Lugt N.M., Acton D., Laird P.W., Linders K., Berns A. Pim-1 levels determine the size of 

early B lymphoid compartments in bone marrow. J Exp Med 178:1665–1673, 1993. PMCID: PMC2191259. 

• van der Lugt N.M., Domen J., Linders K., van Roon M., Robanus-Maandag E., te Riele H., van der Valk 

M., Deschamps J., Sofroniew M., van Lohuizen M., Berns A. Posterior transformation, neurological 

abnormalities, and severe hematopoietic defects in mice with a targeted deletion of the bmi-1 protooncogene. Genes Dev 8:757–769, 1994. PMID: 7926765. 

• Kondo M., Akashi K., Domen J., Sugamura K., Weissman I.L. Bcl-2 rescues T lymphopoiesis, but not B or 

NK cell development, in common- chain-deficient mice. Immunity 7:155–162, 1997. PMID: 9252128. 

• De Vivo I., Cui X., Domen J., Cleary M.L. Growth stimulation of primary B cell precursors by the antiphosphatase Sbf1. Proc Natl Acad Sci U S A 95:9471–9476, 1998. PMCID: PMC21362. 

• Domen J. and Weissman I.L. Hematopoietic stem cells need two signals to prevent apoptosis; BCL-2 can 

provide one of these, Kitl/c-Kit signaling the other. J Exp Med 192:1707–1718, 2000. PMCID: 

PMC2213494. 

Biographical Sketches for each listed Senior/Key Person 2 Page 18

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
• Domen J. and Weissman I.L. HSC and other hematopoietic cells show broad resistance to 

chemotherapeutic agents in vivo when overexpressing BCL-2. Exp Hematol 31:631–639, 2003. PMID: 

12842708. 

• Reya T., Duncan A.W., Ailles L., Domen J., Scherer D., Willert K., Hintz L., Nusse R. and Weissman I.L. A 

role for Wnt signalling in self-renewal of haematopoietic stem cells. Nature 423:409–414, 2003. PMID: 

12717450. 

• Chen B.J., Cui X., Sempowski G.D., Domen J., Chao N.J. Hematopoietic stem cell dose correlates with 

the speed of immune reconstitution after stem cell transplantation. Blood 103:4344–4352, 2004. PMID: 

14976038. 

• Dalal, J., Gandy, K.L. and Domen, J. Role of mesenchymal stem cell therapy in Crohn’s disease. Pediatric 

Res. 71:445-451(2012). PMID: 22430380. 

D. Research Support 

Ongoing Research Support

Katherine B Richardson Endowment Award 7/2011-6/2013

Tolerance induction using myeloid progenitors: Are lymphoid cells essential. 

The aim of this proposal is to test whether tolerance induction by myeloid progenitor cells is dependent on the 

presence of lymphoid cells derived from HSC and multi-potent progenitors present in the MP preparation 

Role: Principal Investigator 

CMH Cancer Research Grant Shreve (PI) 7/2012-6/2014 

Incidence ofVitamin D deficiency in pediatric hematopoietic cell transplant patients. 

The aim of this proposal is to determine to what extend pediatric transplant patients are vitamin D deficient 

before and following transplant, and if that correlates with infection rate and immune function. 

Role: co-investigator 

Completed Research Support

Thoracic Surgery Foundation for Research and Education Gandy (PI) 7/2008–6/2011 

Nina Braunwald Career Advancement Award 

The use of autologous HSC transplantation for tolerance induction. 

The aim of this proposal is the investigate whether resetting the immune system through autologous HCT is a 

viable option to induce tolerance to a transplanted solid organ. 

Role: Co-Investigator

Advancing a Healthier Wisconsin 7/2008–6/2010 

The use of Myeloid Progenitors to improve immune competence after HSC transplantation for tolerance 

induction. 

The aim of this proposal is to investigate the use of third-party myeloid progenitors as an adjuvant therapy to 

HCT as used for tolerance induction for solid organ transplantation. Myeloid Progenitors have the potential to 

reduce the consequences of treatment related neutropenia. 

Role: Principal Investigator 

NIH Small Business Biodefense Programs grant 

1 R43 AI 61856 -01 Fong (PI) 8/2004–7/2006 

Expansion of HSC for rescue in biodefense applications. 

The aim of this proposal is to produce a pre-clinical system based on Cellerant’s proprietary HSC expansion 

technology. This technology entails the transient genetic modification of purified HSC by transduction to 

express genes that result in their ex vivo expansion. 

Role: Co-Investigator 

NIH SBIR grant 

1 R43 AI64156-01 6/2005–6/2006 

Application of expanded progenitors against infections. 

The aim is to characterize and define human myeloid progenitors in various source tissues, and establish 

parameters under which murine myeloid progenitors, generated from highly purified HSC in culture, can be 

used to protect neutropenic mice from potentially lethal infections, e.g. by fungus. The end goal is to develop 

human myeloid progenitors as an off-the-shelf product that can be used to treat neutropenic humans. 

Role: Principal Investigator

Biographical Sketches for each listed Senior/Key Person 2 Page 19

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Medical College of Wisconsin Institutional Research Grant Gandy (PI) 1/2008–1/2009 

The use of autologous HSC transplantation for tolerance induction. 

The aim of this proposal is the investigate whether resetting the immune system through autologous HCT is a 

viable option to induce tolerance to a transplanted solid organ. 

Role: Co-Investigator 

Children’s Research Institute Pilot Innovative Research Award 3/2008–2/2009 

The use of bone marrow to reduce damage due to cardiomyopathy. 

The aim of this proposal is to rigorously test for engraftment by bone marrow derived cells in the myocardium 

of a mouse model of congenital cardiomyopathy. In this model i.v. administration of bone marrow improves 

heart function without readily apparent engraftment. 

Role: Principal Investigator 

CTSI-Medical College of Wisconsin Gandy (PI) 8/2008–10/2009 

The immune competence of children after congenital heart surgery. 

The aim of this proposal is to evaluate the immune function of children in the first year after congenital heart 

surgery to determine if there are detectable immune deficits which changes in practice may improve. 

Role: Co-Investigator

Biographical Sketches for each listed Senior/Key Person 2 Page 20

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
* Last Name

RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 1

* Budget Type:

Enter name of Organization:

* Start Date: * End Date:

* Project Role Base Salary ($)

* Fringe 

Benefits ($) * Funds Requested ($)

9.

8.

7.

6.

5.

4.

3.

2.

1.

Total Funds requested for all Senior Key Persons in the attached file

Total Senior/Key Person

Additional Senior Key Persons:

B. Other Personnel

A. Senior/Key Person

* Number of 

Personnel * Project Role

Acad. 

Months

Sum. 

Months

* Requested 

Salary ($)

Cal. 

Months

Post Doctoral Associates

Graduate Students

Undergraduate Students

Secretarial/Clerical

Total Number Other Personnel Total Other Personnel

Total Salary, Wages and Fringe Benefits (A+B)

Prefix * First Name Middle Name Suffix

* Fringe 

Benefits ($) * Funds Requested ($)

Acad. 

Months

Sum. 

Months

* Requested 

Salary ($)

Cal. 

Months

RESEARCH & RELATED Budget {A-B} (Funds Requested)

* ORGANIZATIONAL DUNS:

OMB Number: 4040-0001 

Expiration Date: 06/30/2011

Project Subaward/Consortium

1

Delete Entry

Dr. Holger Karsunky PD/PI 12.00

1 Research Associate 12.00

Cellerant Therapeutics, inc.

06/01/2013 05/31/2014 Budget Period 

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Detailed Budget - Year 1 Page 21

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
C. Equipment Description

List items and dollar amount for each item exceeding $5,000

Equipment item * Funds Requested ($)

1.

2.

3.

4.

5.

6.

7.

8.

9.

10.

11. Total funds requested for all equipment listed in the attached file

D. Travel

1. Domestic Travel Costs ( Incl. Canada, Mexico and U.S. Possessions)

2. Foreign Travel Costs

Total Travel Cost

Total Equipment

E. Participant/Trainee Support Costs

1. Tuition/Fees/Health Insurance

2. Stipends

3. Travel

4. Subsistence

5. Other

Number of Participants/Trainees Total Participant/Trainee Support Costs

Funds Requested ($)

Funds Requested ($)

RESEARCH & RELATED Budget {C-E} (Funds Requested)

RESEARCH & RELATED BUDGET - SECTION C, D, & E, BUDGET PERIOD 1

* Budget Type:

Enter name of Organization:

* Start Date: * End Date:

Additional Equipment:

* ORGANIZATIONAL DUNS:

Project Subaward/Consortium

06/01/2013 05/31/2014

Cellerant Therapeutics, inc.

Add Attachment Delete Attachment View Attachment

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Detailed Budget - Year 1 Page 22

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
F. Other Direct Costs

1. Materials and Supplies

2. Publication Costs

3. Consultant Services

4. ADP/Computer Services

5. Subawards/Consortium/Contractual Costs

6. Equipment or Facility Rental/User Fees

7. Alterations and Renovations

8.

9.

10.

Total Other Direct Costs

G. Direct Costs

Total Direct Costs (A thru F)

H. Indirect Costs Indirect Cost 

Rate (%)

Indirect Cost 

Base ($)

1.

2.

3.

4.

Cognizant Federal Agency

I. Total Direct and Indirect Costs

Total Direct and Indirect Institutional Costs (G + H)

J. Fee

K. * Budget Justification

Indirect Cost Type

Funds Requested ($)

Funds Requested ($)

* Funds Requested ($)

Funds Requested ($)

Funds Requested ($)

RESEARCH & RELATED Budget {F-K} (Funds Requested)

Total Indirect Costs

(Only attach one file.)

(Agency Name, POC Name, and POC Phone Number)

HHS/ASPR/ACMG Ethan Mueller 

1240-Budget Justification Cellerant.pd

General and Aministrative

RESEARCH & RELATED BUDGET - SECTION F-K, BUDGET PERIOD 1

* Budget Type:

Enter name of Organization:

* Start Date: * End Date:

* ORGANIZATIONAL DUNS:

Project Subaward/Consortium

06/01/2013 05/31/2014

Cellerant Therapeutics, inc.

Budget Period 1

Add Attachment Delete Attachment View Attachment

Next Period

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Detailed Budget - Year 1 Page 23

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
BUDGET JUSTIFICATION 

For Cellerant Therapeutics (Laboratory of Dr. Karsunky) 

Cellerant will use its laboratory resources in San Carlos, CA to accomplish the aims of this grant 

application. The assumptions for the budget have been prepared based on previous experience and 

prior purchases. 

Section A – Senior/Key Person: Dr. Holger Karsunky (PD/PI) will lead and be responsible for the 

scientific management of the project. Dr. Karsunky will contribute his expertise in all aspects of 

myeloid progenitor biology, hematopoietic stem cell biology, in vivo animal models and cell 

therapeutic drug development. 

Name Project Role Proposed % 

effort

% effort on existing grants for 

the proposed budget period

Holger Karsunky PD/PI 10% 90% 

Section B – Other Personnel: We budgeted for 50% time effort for a Research Associate who will 

contribute to the project by harvesting tissues, isolating HSC from mice, ex vivo generation of mouse 

MPC, flow cytometric analysis and sorting, colony formation assays, and cryopreservation. 

Name Project Role Proposed % 

effort

% effort on existing grants for 

the proposed budget period

Greg Boucher RA 50% 50%

Fringe benefits, which are 38.94% of salary requested, cover payroll taxes, employee fringe benefits, 

and workers compensation insurance. 

Section C – Equipment: This proposal includes no equipment budget. All equipment needed is 

currently operational at Cellerant. 

Section D – Travel: We are requesting $ for two trips to the academic partner in Kansas City, 

MO at the beginning of the project and after 6-9 months. Costs are based on an average nonrefundable economy class ticket, two nights per trip at the GSA lodging rate and GSA per diems rates. 

Section F.1 – Materials and Supplies: We are requesting a total of $ for materials and 

supplies. The major cost factors is the purchase of tissue culture reagents and consumables in 

particular recombinant growth factors but also media, media supplements, and culture vessels for the 

ex vivo expansion of mouse myeloid progenitor cells. A second major cost factor are the various 

fluorochrome labeled antibodies needed for flow cytometry to sort the starting material and 

characterize the end product for its compositions. Also included are the costs of procuring up to 540 

mice form various mouse strains that are required to isolate the starting material for the mouse MP. 

Cellerant has three qualified mouse vendors (JAX Mice Services, Charles River Laboratories and 

Taconic) and will also use in-house bred animals. The remaining funds are for general laboratory 

supply. 

Section F.3 – Consultant: No consulting costs are requested as part of this proposal. 

Section F.5 – Subawards/Consortium: The budget for the academic partner is $ ($ 

direct costs plus $ indirect costs). Their budget justification (see below) and their detailed 

budget are provided. 

Budget Justification Page 24

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Section H – Indirect Costs: The G&A rate, currently at %, is used to support all of the 

administrative, operational and contracting matters for Cellerant. Indirect support services include 

vivarium operations, lab infrastructure and facilities, IT support, facilities support, accounting, 

procurement of materials and human resources. The current rate is based upon a bilateral rate 

agreement date Nov 6, 2012 between Cellerant Therapeutics, Inc and HHS/ASPR/ACMG. The 

indirect rates are reviewed annually by an auditor from HHS/ASPR/ACMG and the Peninsula Branch 

office is the Cognizant DCAA office. 

Budget Justification Page 25

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Section A, Senior/Key Person

Section C, Equipment

RESEARCH & RELATED BUDGET - Cumulative Budget

Section D, Travel

Domestic

Section E, Participant/Trainee Support Costs

Foreign

Tuition/Fees/Health Insurance

Stipends

Travel

Subsistence

Other

Number of Participants/Trainees

1.

2.

3.

4.

5.

6.

1.

2.

Section F, Other Direct Costs

1. Materials and Supplies

2. Publication Costs

3. Consultant Services

4. ADP/Computer Services

5. Subawards/Consortium/Contractual Costs

6. Equipment or Facility Rental/User Fees

7. Alterations and Renovations

8.

9.

10.

Totals ($)

Total Number Other Personnel

Total Salary, Wages and Fringe Benefits (A+B)

Other 1

Other 2

Other 3

Section B, Other Personnel

Section J, Fee

Section I, Total Direct and Indirect Costs (G + H)

Section H, Indirect Costs

Section G, Direct Costs (A thru F)

1

	


 		

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Cumulative Budget Page 26

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 1

* ORGANIZATIONAL DUNS:

* Budget Type: ❍ Project ● Subaward/Consortium

Enter name of Organization: Children's Mercy Hospital and Clinics

* Start Date: 06-01-2013 * End Date: 05-31-2014 Budget Period: 1

A. Senior/Key Person

Prefix * First Name Middle Name * Last Name Suffix * Project Role Base Salary

($)

Cal.

Months

Acad.

Months

Sum.

Months

* Requested

Salary ($)

* Fringe

Benefits ($)

* Funds Requested ($)

1. Jos Domen PD/PI 12.00

Total Funds Requested for all Senior Key Persons in the attached file

Additional Senior Key Persons: File Name: Mime Type: Total Senior/Key Person

B. Other Personnel

* Number of

Personnel

* Project Role Cal.

Months

Acad.

Months

Sum.

Months

* Requested

Salary ($)

* Fringe

Benefits

* Funds Requested

($)

Post Doctoral Associates

Graduate Students

Undergraduate Students

Secretarial/Clerical

1 Research Technician 12.00

1 Research Technician 12.00

2 Total Number Other Personnel Total Other Personnel

Total Salary, Wages and Fringe Benefits (A+B)

RESEARCH & RELATED Budget {A-B} (Funds Requested)

Tracking Number: GRANT11279532 OMB Number: 4040-0001

Expiration Date: 06/30/2011

Subaward 1 Page 27

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
RESEARCH & RELATED BUDGET - SECTION C, D, & E, BUDGET PERIOD 1

* ORGANIZATIONAL DUNS:

* Budget Type: ❍ Project ● Subaward/Consortium

Enter name of Organization: Children's Mercy Hospital and Clinics

* Start Date: 06-01-2013 * End Date: 05-31-2014 Budget Period: 1

C. Equipment Description

List items and dollar amount for each item exceeding $5,000

Equipment Item * Funds Requested ($)

Total funds requested for all equipment listed in the attached file

Total Equipment

Additional Equipment: File Name: Mime Type:

D. Travel Funds Requested ($)

1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions)

2. Foreign Travel Costs

Total Travel Cost

E. Participant/Trainee Support Costs Funds Requested ($)

1. Tuition/Fees/Health Insurance

2. Stipends

3. Travel

4. Subsistence

5. Other:

Number of Participants/Trainees Total Participant/Trainee Support Costs

RESEARCH & RELATED Budget {C-E} (Funds Requested)

Tracking Number: GRANT11279532 OMB Number: 4040-0001

Expiration Date: 06/30/2011

Subaward 1 Page 28

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
RESEARCH & RELATED BUDGET - SECTIONS F-K, BUDGET PERIOD 1

* ORGANIZATIONAL DUNS:

* Budget Type: ❍ Project ● Subaward/Consortium

Enter name of Organization: Children's Mercy Hospital and Clinics

* Start Date: 06-01-2013 * End Date: 05-31-2014 Budget Period: 1

F. Other Direct Costs Funds Requested ($)

1. Materials and Supplies

2. Publication Costs

3. Consultant Services

4. ADP/Computer Services

5. Subawards/Consortium/Contractual Costs

6. Equipment or Facility Rental/User Fees

7. Alterations and Renovations

8. Mouse Costs Aim 1

9. Mouse Costs Aim 2

Total Other Direct Costs

G. Direct Costs Funds Requested ($)

Total Direct Costs (A thru F)

H. Indirect Costs

Indirect Cost Type Indirect Cost Rate (%) Indirect Cost Base ($) * Funds Requested ($)

1. General and Administrative

Total Indirect Costs

Cognizant Federal Agency

(Agency Name, POC Name, and POC Phone Number)

I. Total Direct and Indirect Costs Funds Requested ($)

Total Direct and Indirect Institutional Costs (G + H)

J. Fee Funds Requested ($)

K. * Budget Justification File Name: 1243-Budget Justification Domen.pdf Mime Type: application/pdf

(Only attach one file.)

RESEARCH & RELATED Budget {F-K} (Funds Requested)

Tracking Number: GRANT11279532 OMB Number: 4040-0001

Expiration Date: 06/30/2011

Subaward 1 Page 29

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
RESEARCH & RELATED BUDGET - Cumulative Budget

Totals ($)

Section A, Senior/Key Person

Section B, Other Personnel

Total Number Other Personnel 2

Total Salary, Wages and Fringe Benefits (A+B)

Section C, Equipment

Section D, Travel

1. Domestic

2. Foreign

Section E, Participant/Trainee Support Costs

1. Tuition/Fees/Health Insurance

2. Stipends

3. Travel

4. Subsistence

5. Other

6. Number of Participants/Trainees

Section F, Other Direct Costs

1. Materials and Supplies

2. Publication Costs

3. Consultant Services

4. ADP/Computer Services

5. Subawards/Consortium/Contractual Costs

6. Equipment or Facility Rental/User Fees

7. Alterations and Renovations

8. Other 1

9. Other 2

10. Other 3

Section G, Direct Costs (A thru F)

Section H, Indirect Costs

Section I, Total Direct and Indirect Costs (G + H)

Section J, Fee

Tracking Number: GRANT11279532 OMB Number: 4040-0001

Expiration Date: 06/30/2011

Subaward 1 Page 30

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
BUDGET JUSTIFICATION 

For Subawardee - Children’s Mercy Hospital/University of Missouri, Kansas City (Laboratory of 

Dr. Domen) 

Personnel. 

The personnel listed in this proposal is in place and is experienced in performing the assays and 

techniques described in this proposal. 

Jos Domen (10% effort) will function as PI for the efforts at Children’s Mercy Hospital and will 

be mainly involved in designing and interpreting experiments performed by the academic partner. He 

will also be involved in other aspect of the experiments as needed. 

Lei Sun (25% effort) and Yongwu Li (25% effort) both have worked in the lab for years. Both 

are experienced in obtaining bone marrow, staining it with antibodies, sorting HSC and MP, expand 

HSC in culture into MP, placing skin grafts and flow cytometric analysis of mice. Yongwu Li will focus 

on the animal experiments such as placements and follow-up of skin grafts. Lei Sun will focus on cell 

sorting and analysis, HSC to MP cultures and the in vitro assays as described in Aim 1. In addition 

she will be responsible for tasks such as ordering supplies. 

Equipment. N/A

Supplies. 

Both the FACS sorts as well as the flow cytometric analysis require fluorochrome labeled antibodies. 

A typical Hematopoietic Stem Cell sort requires a total of 11 different antibodies in 4 colors, while a 

typical analysis uses 5 to 6 different labeled antibodies. While we have complete sets of antibodies we 

will have to replace regularly as tubes are emptied. We are budgeting for 12-15 replacements per 

year (costs varies with fluorochrome). 

ScilVet Bloodcell counter. In flow cytometric analysis of blood we typically include a blood cell count to 

determine absolute cell numbers. The lab has a ScilVet ABC veterinary blood cell counter for this 

purpose. The requested consumables should allow us to do approx 900 cell counts, each requiring 

12μl of blood. The machine will also be used for cell counts when harvesting and analyzing organs. 

Flow cytometry will be used extensively for analysis purposes. The six-color Attune is available for this 

purpose and doesn’t require an hourly fee. However, we have budgeted for consumables such as 

focusing buffer, wash and shutdown solution and test beads, all of which are essential in using the 

instrument. 

Sort time. We have budgeted for sixteen 1 hour sorts at the KU Medical Center Flow Core facility. 

This should allow us to sort the HSC needed in these experiments. MP will be provided by Cellerant 

Therapeutics. In addition to HSC we may sort MP subpopulations for the MLR assays in Aim 1. 

Histology. The outcome of skin graft experiments typically is very clear by gross morphology, but 

trachea grafts need to be evaluated microscopically. We have budgeted for histological analysis of 

150 samples, using the core facility at Children’s Mercy Hospital. The procedures include embedding, 

sectioning, H&E stain as well as two immune stains. We aim to use immunohistochemistry to test for 

graft infiltrating lymphocytes or MP-derived cells in tissues. 

Other consumables. In addition to these major categories there will be need for a variety of 

consumables, including plasticware (FACS tubes, tissue culture plates), media and buffers, IgG for 

blocking, surgical supplies such as sutures and anesthesia. 

Travel. Travel costs will be used for a meeting with the corporate partner in San Carlos, CA. This in 

addition to the meetings listed under the Cellerant budget justification section. In addition, travel funds 

may be used to present the data in a meeting such as the annual meeting of the American Society of 

Hematology or the International Society for Heart and Lung Transplantation. 

Subaward 1 Budget Justification Attachment Page 31

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Patient Care. N/A 

Other. 

Mouse purchasing. Typical experiments require 50 mice (15 hosts, plus 5 HSC-donors, and upto 30 

trachea or skin graft donors). We expect to use approx. 700 mice, at least 6 groups for aim 1, and 8 

groups for aim 2. Mouse purchase prices average $20 for young mice of these standard strains. 

Mouse per diems. Host mice, 3 cages per group, will be kept one to six months to evaluate 

hematopoietic transplantation and trachea or skin graft. Trachea grafts are harvested early (one 

month) while skin grafts are followed longer. Donor mice will be kept for an average less than one 

month before tissue harvest. We expect that mice, on average, will be kept for 2 months. Cage cost is 

$0.54 per day and we expect approximately 188 cages for the experiments described. 

Subaward 1 Budget Justification Attachment Page 32

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
SBIR/STTR Information OMB Number: 4040-0001 

Expiration date: 06/30/2011 

* Program Type (select only one)

* SBIR/STTR Type (select only one)

* 1a. Do you certify that at the time of award your organization will meet the eligibility criteria for a small business as defined in the funding 

opportunity announcement?

* 2. Does this application include subcontracts with Federal laboratories or any other Federal Government agencies?

* If yes, insert the names of the Federal laboratories/agencies:

* 3. Are you located in a HUBZone? To find out if your business is in a HUBZone, use the mapping utility provided by the Small Business 

* 4. Will all research and development on the project be performed in its entirety in the United States?

* Explanation:

* 5. Has the applicant and/or Program Director/Principal Investigator submitted proposals for essentially equivalent work under other 

Federal program solicitations or received other Federal awards for essentially equivalent work? 

* If yes, insert the names of the other Federal agencies:

* 6. Disclosure Permission Statement: If this application does not result in an award, is the Government permitted to disclose the title of 

your proposed project, and the name, address, telephone number and e-mail address of the official signing for the applicant organization, 

to organizations that may be interested in contacting you for further information (e.g., possible collaborations, investment)?

* 7. Commercialization Plan: If you are submitting a Phase II or Phase I/Phase II Fast-Track Application, include a 

Commercialization Plan in accordance with the agency announcement and/or agency-specific instructions.

* Attach File:

Questions 1-7 must be completed by all SBIR and STTR Applicants:

If no, provide an explanation in an attached file.

Administration at its web site: http://www.sba.gov 

SBIR STTR

Both (See agency-specific instructions to determine whether a particular agency allows a single submission for both SBIR and STTR)

Phase I Phase II

Fast-Track (See agency-specific instructions to determine whether a particular agency participates in Fast-Track)

Yes

No

65

Yes

No

Yes

No

Yes

No

Yes

No

Yes

No

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* 1b. Anticipated Number of personnel to be employed at your organization at the time of award.

	


 		

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SBIR/STTR Information Page 33

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
* 8. Have you received SBIR Phase II awards from the Federal Government? If yes, provide a company commercialization history in 

accordance with agency-specific instructions using this attachment. 

* Attach File:

* 9. Will the Project Director/Principal Investigator have his/her primary employment with the small business at the time of award?

* 10. Please indicate whether the answer to BOTH of the following questions is TRUE:

(1) Does the Project Director/Principal Investigator have a formal appointment or commitment either with the small business directly 

(as an employee or a contractor) OR as an employee of the Research Institution, which in turn has made a commitment to the 

small business through the STTR application process; AND 

(2) Will the Project Director/Principal Investigator devote at least 10% effort to the proposed project?

* 11. In the joint research and development proposed in this project, does the small business perform at least 40% of the work and the research 

institution named in the application perform at least 30% of the work?

SBIR-Specific Questions:

STTR-Specific Questions:

SBIR/STTR Information

Questions 8 and 9 apply only to SBIR applications. If you are submitting ONLY an STTR application, leave questions 8 and 9 blank and proceed to 

question 10.

Questions 10 and 11 apply only to STTR applications. If you are submitting ONLY an SBIR application, leave questions 10 and 11 blank.

Add Attachment Delete Attachment View Attachment

Yes

No

Yes

No

Yes

No

Yes

No

	


 		

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SBIR/STTR Information Page 34

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
PHS 398 Cover Page Supplement

1. Project Director / Principal Investigator (PD/PI)

* Title:

* Street1:

* City:

* Country:

Street2:

County/Parish:

* State:

* Zip / Postal Code:

Prefix: * First Name:

Middle Name:

* Last Name:

Suffix:

2. Human Subjects

Clinical Trial?

* Agency-Defined Phase III Clinical Trial?

* First Name:

Middle Name:

* Last Name:

Suffix:

3. Applicant Organization Contact

Person to be contacted on matters involving this application

* Phone Number: Fax Number:

Email:

OMB Number: 0925-0001 

Province:

Prefix:

Dr. Holger

Karsunky

No Yes

No Yes

Dr. Holger

Karsunky

Director, Preclinical Development

1561 Industrial Road

San Carlos

USA: UNITED STATES

CA: California

94070-4111

	


 		

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Clinical Trial & HESC Page 35

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
4. Human Embryonic Stem Cells

* Does the proposed project involve human embryonic stem cells?

If the proposed project involves human embryonic stem cells, list below the registration number of the 

specific cell line(s) from the following list: http://stemcells.nih.gov/research/registry/. Or, if a specific 

stem cell line cannot be referenced at this time, please check the box indicating that one from the 

registry will be used: 

Specific stem cell line cannot be referenced at this time. One from the registry will be used.

PHS 398 Cover Page Supplement

No Yes

Cell Line(s):

	


 		

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Clinical Trial & HESC Page 36

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
PHS 398 Research Plan

1. Application Type:

From SF 424 (R&R) Cover Page. The response provided on that page, regarding the type of application being submitted, is repeated for your 

reference, as you attach the appropriate sections of the Research Plan.

*Type of Application:

OMB Number: 0925-0001 

2. Specific Aims

3. *Research Strategy

6. Protection of Human Subjects

7. Inclusion of Women and Minorities

8. Targeted/Planned Enrollment Table

9. Inclusion of Children

10. Vertebrate Animals

13. Consortium/Contractual Arrangements

14. Letters of Support

15. Resource Sharing Plan(s)

16. Appendix

1. Introduction to Application 

(for RESUBMISSION or REVISION only)

Human Subjects Sections

2. Research Plan Attachments: 

 Please attach applicable sections of the research plan, below.

Other Research Plan Sections

5. Progress Report Publication List

11. Select Agent Research

12. Multiple PD/PI Leadership Plan

4. Inclusion Enrollment Report

New Resubmission Renewal Continuation Revision

Add Attachment Delete Attachment View Attachment

1241-SpecificAimsSTTR2012v2 Add Attachment Delete Attachment View Attachment

1242-ResProposalSTTR2012v20 Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

1246-Combined Vertebrate An Add Attachment Delete Attachment View Attachment

1247-11 SelectAgentResearch Add Attachment Delete Attachment View Attachment

1248-12 MultiplePDPI_FINAL.p Add Attachment Delete Attachment View Attachment

1249-STTR Letter verifying e Add Attachment Delete Attachment View Attachment

Add Attachment Delete Attachment View Attachment

1250-15 ResourceDataSharing Add Attachment Delete Attachment View Attachment

Add Attachments Remove Attachments View Attachments

 


  		

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List of Research Plan Attachments Page 37

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
SPECIFIC AIMS

Despite many advances in the field of transplantation, 10-year graft survival rates are still low: 20% for lung 

transplantation and just over 50% for heart transplantation. The latter has one of the highest long-term success 

rates. Rejection is thought to cause the majority of graft loss. Operational tolerance, the ability of an otherwise 

immunocompetent host to accept a transplanted organ without immunosuppression, would likely improve longterm organ function and offer significant benefits for recipient’s quality of life and longevity. Specifically for lung 

transplantation, the most important cause of late mortality is obliterative bronchiolitis, characterized by specific

lesions and a decrease in lung function [1]. Despite decades of research, no widely applicable clinical protocol 

has emerged that can achieve consistent operational tolerance. Certain bone marrow transplantation regimens 

are capable of tolerance induction, but widespread clinical application has been limited. Important limitations 

are the severity of the necessary preconditioning regimens, with its associated morbidity and mortality

(including severe immuno-incompetence) and the availability of adequate bone marrow grafts. We have 

recently identified a population of myeloid progenitor (MP) cells that may allow the development of clinical 

protocols that will overcome these hurdles. Cellerant Therapeutics is a company that has developed methods 

to obtain and test mouse and clinical grade human MP [2]. Research by the academic partner, currently at 

Children’s Mercy Hospital/University of Missouri Kansas City, has shown that injection of mouse MP cells and 

purified hematopoietic stem cells (HSCs) into irradiated hosts leads to long-term, MP donor-specific tolerance 

[3]. 

The Domen lab continues to expand on these promising observations in important ways. (1) We have 

demonstrated that MP cells purified from bone marrow and MP cells expanded in short-term cultures induce 

specific tolerance when infused into irradiated recipients. (2) MP cells function in the context of both allogeneic 

and autologous HSC transplantation. (3) Tolerance is induced when the graft is either placed at the time of MP 

infusion or at a later time. (4) Myeloid cells are sufficient for tolerance induction. MP cells from Rag2 -/-

Il2rg -/-

mice, incapable of producing functional lymphocytes because they lack the Rag2 recombinase and signaling 

through many Interleukin receptors, prove that the myeloid cells alone are sufficient. (5) A minor degree of 

mismatching between MP and graft seems to be acceptable. Based on these observations, Cellerant 

Therapeutics and Children’s Mercy/Domen Lab propose to jointly investigate the following hypotheses:

(I) One or more, organ graft-matched, myeloid progenitor-derived cell populations are essential and sufficient 

for cellular tolerance induction in multiple transplant models, including lung transplant.

(II) MP cells can induce tolerance for both fully matched and partially matched allogeneic grafts. 

Specific aims

1. Aim: Show that MPc can be used to induce tolerance for lung transplants in a mouse model using 

heterotopic tracheal allografts. 

Milestone: No luminal occlusion and maintenance of epithelium in allogeneic (MP-matched) tracheal 

transplants.

2. Aim: Determine the degree of mismatching allowed for tolerance induction by MP. 

Milestone: Determine transplant acceptance rates induced with haploidentical MP and MP matched for 

major but not minor MHC. 

Significance. The long-term survival of lung transplant recipients (1,200 to 1,400 transplants per year) is low. 

As such this is a patient population that could benefit greatly from an efficient tolerance induction regimen. The 

high failure rate in these patients will limit both the size and time of the clinical trials needed to show efficacy. 

Our research has defined, in a preclinical model, a novel tolerance induction protocol. Our proposed research 

will focus on testing the efficacy of this approach in lung transplantation models. Human MP cells, CLT-008,

are currently undergoing clinical trials in umbilical cord blood transplantation and high dose chemotherapy 

leukemia trials, both aimed at reducing infectious complications. Our efforts in this project will set the stage to 

translate this preclinical approach, tolerance induction by MP, into a testable and clinically relevant product. 

Our goal is to enable the prompt integration of MP cells into transplantation strategies, starting with organ 

systems that currently enjoy limited long-term success, such as lung transplantation. 

Innovation. Few, if any, translational research protocols aimed at tolerance induction focus on the use of 

clinically available MP cells. This novel approach, specifically aimed at defining the components of a protocol 

that has the potential for broad clinical application, will lead to a clinical paradigm shift once translated into an 

approved and commercially available clinical option.

Specific Aims Page 38

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
RESEARCH STRATEGY

(A) SIGNIFICANCE

Short-term survival following cardiopulmonary organ transplant is excellent but long-term survival remains 

limited. Only 20% of the transplanted lungs remain functional ten years after transplantation [4], even with 

continued immunosuppression. Daily life is complicated by the staggering morbidity associated with the

diagnosis and treatment of frequent and debilitating rejection episodes. Long-term immunosuppression, 

essential to delay rejection, can itself be considered a complication, since the associated morbidities are 

numerous and debilitating (including neural and renal toxicity and higher risk for cancer and infections [5-8]). 

Operational tolerance, the acceptance of the transplanted organ into an otherwise immunocompetent 

host, should greatly improve long-term outcome, improve patient quality of life and reduce the economic 

burden on society. However, tolerance induction remains an elusive goal [9]. Tolerance will especially benefit 

pediatric transplant recipients, as transplanted organs need to function for six or more decades to restore a 

normal lifespan in children. Many approaches to tolerance induction have been tested (reviewed in [10-20]). 

Table 1 below categorizes some of these treatment options and their limitations.

Immunosuppressant (standard therapy, no tolerance induction) Effective short term, less so long term. Significant side effects.

Costimulatory blockade, T cell depletion Limited effectiveness alone, may be good in combination therapy.

Hematopoietic (stem) Cell Transplantation, BMT Very effective, works at multiple levels, but severe co-morbidity. 

Progenitor Cell Transplantation (Myeloid Progenitors) This proposal. Effective. Can work through different cell types.

Other cellular therapies (eg Treg, macrophages or dendritic cells) Promising but addresses fewer mechanisms (single cell type).

Table 1. Comparison of standard therapy and several tolerance inducing strategies.

The success of hematopoietic cell transplantation (HCT) has been well established with various cell

populations, including rigorously purified hematopoietic stem cells (HSCs) [21]. However, despite decades of 

work clinical translation has been limited to small-scale trials [12, 18, 19, 22-24], mainly due to HCT-associated 

complications. Although HCT-related outcomes continue to improve incrementally [25], increased acceptance 

and use of HCT for treatment of non-malignancies will require significant reductions in morbidity and mortality. 

The power of myeloid cells in tolerance induction has been more fully appreciated recently. This 

proposal is centered on a population of cells called myeloid progenitor (MP) cells that give rise to all cells of the 

myeloid lineage. Key characteristics of myeloid progenitors include: clinically availability; minimal to no 

side effects in ongoing clinical trials and importantly; specific tolerance induction potential in our 

experimental model system. Our proposal focuses on obtaining the data needed to bring this work to 

commercial development and clinical availability in the near future.

Cellerant Therapeutics, Inc has worked on developing MP as an off the shelf clinical product (product 

name CLT-008) for the past 8 years, and is currently testing CLT-008 in two clinical trials. CLT-008 is being 

developed for two indications: (1) To prevent infectious complications in severely neutropenic patients and (2) 

to treat acute radiation syndrome (ARS). This proposal, based on laboratory work performed by the Domen lab 

at the Medical College of Wisconsin, Milwaukee, WI and Children’s Mercy Hospital in Kansas City, MO, aims to 

add tolerance induction for solid organ transplantation as an indication for the use of MP. This will serve a 

currently unmet medical need. However, as tolerance induction requires matched MPs, the requirements for 

production will differ to some extend from those in the current clinical programs.

Development of CLT-008 as a marketable product is well underway, including ongoing clinical trials and 

scaling up of production, the latter aided by Contract HHSO 100201000051C with the Biomedical Advanced 

Research and Development Authority (BARDA) to develop and stockpile CLT-008 for the treatment of ARS. 

This should greatly aid in the eventual commercial development of CLT-008 for tolerance induction, even if 

modifications in the production process are found to be necessary for use as discussed in this proposal. 

(B) INNOVATION

Our proposal represents a major change to transplantation strategies. Our investigation centers on the unique 

properties of myeloid progenitor cells as inducers of tolerance. This approach includes a dramatic shift away 

from the currently explored cellular therapy strategies that require the use of HSC-containing populations, 

lymphocyte populations, or mature myeloid populations, most of which require the creation of stable chimeras. 

We have characterized MP cells and shown that they induce antigen-specific tolerance that permits animals to 

accept skin grafts without the need for sustained high level hematopoietic chimerism and its associated 

complications [3]. MP cells give rise to all cells of the myeloid lineage and recent data demonstrates the role of 

multiple myeloid populations in tolerance induction. By using progenitor cells that can differentiate into many 

potential effector populations, the potential of all of these populations will be harnessed. However, to translate 

this into clinical practice we will need to define a path. By combining the academic preclinical expertise with 

Cellerant’s expertise in clinical use of MP we expect to be able to define a path such that clinical trials for this 

innovative approach will be feasible at the end of this proposal.

Research Strategy Page 39

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
(C) APPROACH

Clinical Use of MP cells. Cell surface marker combinations 

identifying MP were first characterized in the lab of Irving 

Weissman [26, 27]. Effectiveness of purified mouse MP 

against infections was first demonstrated in Dr. Janice 

Brownʼs lab at Stanford University [28, 29]. Subsequently, 

human MP were developed for clinical use by Cellerant 

Therapeutics [30]. MP are currently being tested in patients 

with hematologic malignancies receiving umbilical cord blood 

transplantation or chemotherapy to protect against infections. 

They can be cryopreserved, do not cause graft-versus-host 

disease, and can be used without HLA matching [31, 32]. MP 

give rise to a variety of cells that have been implicated in 

tolerance induction. MP comprise a set of progenitors that 

can generate all myelo-erythroid lineages [26, 27], including 

dendritic cells (DC) [33] and myeloid-derived suppressor cells 

(MDSC). Myeloid cells comprise a heterogeneous cell 

compartment that is far more complex and compartmentalized 

in pro-inflammatory and immunosuppressive cells than has 

been realized [34]. MDSC and Tolerance. MDSC have been 

mostly studied in the context of cancer, but accumulating evidence indicates an important role in tolerance 

induction [35-37]. Models studied include a rat kidney transplant model [38] in which MDSC are essential but 

not sufficient. In a heart transplantation model in mice suppressive monocytes (CD11c+

, CD11b+, CD115+) are 

essential for tolerance and their depletion prevented tolerance. Transfer of monocyte/dendritic cell (DC) 

precursors, but not DC precursors rescued the tolerance inducing phenotype [39]. Co-transplantation of MDSC, 

but not DC precursors, with islet allografts has been reported to results in allograft tolerance via B7-H1-

mediated enhancement of Tregs [40, 41]. DC and Tolerance. It has long been known that DCs can induce 

antigen-specific tolerance [42-46]. Donor-derived DCs prolong graft acceptance [47], and when combined with 

co-stimulatory blockade or total lymphoid irradiation, induce lasting specific tolerance [48, 49]. In some cases, 

tolerance induced by donor DCs has been reported to involve delivery of donor antigen to recipient antigen 

presenting cells [50]. Donor-derived DCs induce specific tolerance when given simultaneously with the organ 

graft [51]. Tolerance and sublethal preconditioning protocols. Over the last decade sublethal 

preconditioning protocols for HCT and tolerance induction have been developed. The Boston group led by 

Sykes, Sachs, and Spitzer uses cyclophosphamide, thymic irradiation and lymphocyte depleting antibodies for 

preconditioning [22, 52]. Trials have been in effect for a decade. Though tolerance can be induced, the protocol 

has failed in sensitized individuals or donor/recipient pairing of 

more than haplo disparity. A protocol centered around total 

lymphoid irradiation and anti-thymocyte globulin (ATG), has 

been developed by Strober and colleagues [53]. Tolerance 

induction is limited by recipient/donor genetic disparity [54]. 

More recently, a trial has been conducted by Ildstad and 

Leventhal [24] using facilitator cells with which we also have 

experience [55]. Though this protocol is successful across 

significant HLA disparity, it has failed in sensitized patients and 

the implications of the high-level donor-derived reconstitution 

and resultant prolonged immunoincompetence are still unclear.

Preliminary Studies. Studies at Cellerant Therapeutics have 

established that MP, derived from ex vivo expanded 

progenitors, are fully functional with respect to their ability to 

reduce susceptibility to infections. The Domen Lab has recently 

published that MP can be used to induce specific tolerance for 

subsequently transplanted skin grafts in mouse [3]. The 

recipient animals were immunocompetent and rejected grafts 

unmatched to host, HSC donor, or MP donor. We have since 

significantly expanded these observations as detailed below. 

Figure 1. Tolerance is achieved even when skin 

grafts are placed simultaneously with HCT.

BALB/c hosts, HSC donor AKR, FVB or C57BL/6 

mice, MP donors FVB or C57BL/6 mice. Left 

overall survival of matched and unmatched grafts, 

right morphology (week 7) and histology (week 12) 

post transplant confirming the presence of tail skin 

at the graft site with its typical thick epidermis [3]. 

Figure 2. Tolerance induction by MP does not 

require graft-matched lymphocytes. Skin graft 

survival in BALB/c mice reconstituted with 4,000 FVB 

HSC and 100,000 B10; B6-Rag2-/-Il2rg-/- MP. Dotted 

line: AKR skin grafts, unmatched to host, HSC or MP 

donor. Solid lines, matched grafts: no graft failures in 

this data set. Skin grafts placed simultaneous with the 

transplantation of the hematopoietic cells. Grafts 

were followed up to 200 days.

Research Strategy Page 40

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
✱ Skin grafts can be placed at the time of MP 

administration. Our original model of performing the skin 

grafts two months after HCT confines clinical translation to 

transplants from living donors. We have determined that 

specific tolerance can be obtained when the skin graft is 

placed at the same time as the MP graft, broadening the 

applicability of this strategy to deceased donor organs 

(Figs.1-2). (submitted for publication)

✱ Matched myeloid cells are the only essential cells. We 

have demonstrated that tolerance induction only requires 

myeloid cells. B10;B6-Rag2-/-

Il2rg-/- mice lack both the Rag2 recombinase and the gamma chain, essential for signal 

transduction, common to the receptors for IL-2, IL-4, IL-7, 

IL-9, IL-15 and IL-21. As a consequence, they have no Tcells, B cells or NK cells [56]. MP cells from these mice 

induce tolerance for matched skin grafts (Fig. 2), (submitted 

for publication).

✱ Allogeneic stem cell transplantation is not essential.

Allogeneic MP cells induce tolerance when given 

simultaneously with host-autologous, rather than allogeneic, 

HSCs (Fig. 3). Importantly, this indicates that the full 

replacement of the hematopoietic system through an allogeneic HSC transplant is not necessary and that it 

should be possible to develop sublethal regimens for the temporary engraftment of MP. 

✱ Expanded and cryopreserved MP (MPc) are fully functional in our tolerance induction model (Fig. 3). While it 

is possible to sort sufficient quantities of MP from mouse bone marrow, this technique can not generate clinical 

quantities. Short-term expansion in vitro under defined conditions [2, 30, 32] is used for generating clinical MP 

preparations [57]. E.g. in the group with skin grafts placed after two month 10 mice received MP sorted from 

BM, 16 mice MPc expanded from HSC (shown as single group (dashed grey line) in Fig. 3). Skin graft survival 

does not differ (90 vs 81%, p=0.58, logrank) between sorted or ex vivo expanded and cryopreserved MP.

Aim 1: Show that MPc can be used to induce tolerance for lung transplants in a mouse model using 

tracheal allografts. Lung transplant, with its low long-term graft survival rate, is a primary candidate to test 

novel approaches aimed at prolonging the useful lifespan of transplanted organs for several reasons. There is 

the potential to show significant improvements faster and with a more limited set of patients than otherwise 

necessary. In addition, it would help patients whose only alternative currently is a re-transplant, which is not 

always available. The main cause of late failure in lung transplant is bronchiolitis obliterans syndrome (BOS), 

clinically evident as decreased lung function, and morphologically characterized by obliterative bronchiolitis 

(OB). Airway epithelium becomes a target of the alloimmune response, resulting in loss of airway patency. 

Several models have been developed to study OB. In the Phase I part we propose to utilize a tracheal 

transplant, a model developed twenty years ago [58] which is still used extensively to study OB [59-63]. In this 

assay allografts lose their epithelium 

and airway patency following 

transplantation, while isografts retain it. 

Major advantages of this model are the 

ease of use and the reproducibility of 

the assay [64]. We will use this model 

to test whether addition of graft 

matched MP can prevent occlusion and 

loss of airway epithelium in 

transplanted trachea (Fig. 4). The 

readout should be clear and with paired 

analysis (three grafts will be placed per 

mouse) we calculate that groups of 15 

grafts will give us sufficient power (as 

described in the data analysis).

Figure 3. Tolerance can be achieved with graftmatched MP and host-matched HSC. BALB/c hosts 

were reconstituted with 250 BALB/c HSC and 105

MP-directly sorted from BM or 106 MPc-derived in 

ten day culture from HSC (the latter are more 

heterogeneous). Skin grafts were placed at the time of 

HCT (solid black line) or two months later (dashed 

grey): Skin graft survival does not differ (logrank).

Fig 4. Experimental design for Aim 1. BALB/c mice will be lethally irradiated, 

injected with 4,000 FVB HSC and 500,000 C57BL/6 MPc, after which trachea 

grafts are placed (all on same day) from C57BL/6 (MP-matched), FVB (HSCmatched) and AKR (not-matched). Order of trachea grafts will vary randomly. 

Primary outcome will be airway occlusion, secondary loss of epithelium. 

Research Strategy Page 41

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Division of the work. Placement and monitoring of the trachea graft in mice will be performed by the 

academic partner. Sorting of HSC, ex vivo expansion of HSC into MPc and cryopreservation of the MPc will be 

performed by the corporate partner. All cryopreserved MPc will undergo lot release testing that (viability, 

composition, function, endotoxin levels, and bioburden) to ensure a product with consistent purity and potency.

Milestone: Preventing occlusion in mismatched allogeneic tracheal transplants. The goals of this aim will be 

considered met if we can demonstrate in a group of fifteen mice that allogeneic trachea’s have significantly 

reduced luminal occlusion (<50% occlusion by surface area on all cross sections of H&E stained slides)

beyond 30 days post transplant in mice that have been given MP matching the trachea graft prior to or 

simultaneous with the tracheal transplant. Control groups (on the same mice) will include (i) tracheas that do 

not match MP (or HSC or host), these controls should be occluded, and (ii) trachea’s matched to HSC or host, 

these should not occlude. In addition to luminal occlusion we will score loss of epithelium as visible on H&E 

stained sections, as a secondary sign of rejection.

Details of the model. We will use a variation of our established mouse model for the experiments in Aim 1. 

We will lethally irradiate (2x4.25 Gy, 4 hours apart using a RadSource 2000 irradiator at 160 kV, 25 mA, 

calibrated using chemical dosimetry in collaboration with J.D. van Horn, Dept of Chemistry, UMKC) host mice 

(typically BALB/c; other strains require a higher radiation dose). The mice will be injected on the day of 

irradiation with defined combinations of fluorescence-activated cell sorting (FACS)-purified HSCs [3, 55, 65]. 

Depending on the strain of origin (Table 2), one of two phenotypes of HSC will be sorted from BM: for CD90.1 

strains such as FVB we will use 4,000 cells of phenotype CD117+

, CD90.1lo, Lin-, Sca-1+ while from CD90.2 

strains (such as C57BL/6 or BALB/c) we will use 104 cells of phenotype CD117+

, Lin-, Sca-1+. Prior to injection, 

the HSCs will be mixed with either 105

 FACS-purified bone marrow derived MP cells (Linneg/low, CD117+, Sca-1-) 

or 106

 MPc cells expanded from FACS-purified HSCs in a short-term culture (8-10 days in Xvivo15 medium 

supplemented with recombinant mouse SCF, Flt3L, and Tpo) [2]. On the day of injection tracheas are placed 

using published procedures [58, 66]. Donor mice are euthanized and the heart lung block is excised. The 

trachea and main bronchi are removed and implanted in the host mouse in a subcutaneous pocket on the 

back. Typically a mouse will be transplanted with both an MP-matched trachea, an HSC or host-matched 

trachea and an unmatched trachea, placed on either side of the 

midline below the shoulder bone. Trachea’s will be harvested 

from euthanized recipients on days 7, 14 and 30 following 

transplantation and analyzed by (immuno) histochemistry and 

flow cytometry for donor derived cells and status of the trachea 

graft. The most important analysis will be at day 30, as defined 

in milestones. Flow cytometry will focus on detection of 

lymphocytes, either in the graft itself, or systemically in the 

hematopoietic tissues of the host. This analysis will include 

mixed lymphocyte reactions (MLR) aimed at testing 

activation/proliferation of Celltrace Far Red labeled 

lymphocytes by various activators (irradiated host-, donor- or 

third party lymphocytes) to test in vitro for tolerance.

Potential Problems/Alternative Approaches. The tracheal transplant model has been used successfully in a 

large number of studies. The simplicity of surgical techniques and reproducibility of outcome should greatly 

facilitate the generation of data necessary to move this project forward. More sophisticated and complex 

models in the form of orthotopic transplantation of trachea [64], or orthotopic lung transplantation [67, 68], will 

be considered for the second phase of this project if we find questions that the heterotopic trachea transplant 

model can not answer. More important will be adaptations of this model to test human MPc [69]. If acceptance 

of the graft with HSC but not with culture derived MPc is seen, we are prepared to either increase the dose of 

MPc, change their culture conditions, or resort to using freshly purified MP from the bone marrow. 

Aim 2: Determine the degree of mis-matching allowed for tolerance induction by MP. 

Our data, published as well as unpublished, has clearly established that myeloid cells in the MP preparations 

have the potential to induce tolerance for simultaneously or subsequently placed solid organ grafts that are 

fully matched to the MP’s. However, it has not yet been determined whether matching needs to be absolute 

(clinically that means the cells have to be derived from the organ donor) or whether it is sufficient to use MP 

from a donor matching only major histocompatibility antigens. The latter would translate clinically into a 

scenario where MP with known combinations of transplantation antigens can be stockpiled. When organs 

Strain MHC CD90 CD45

AKR k 1 2

BALB/c d 2 2

CBA k 2 2

C57BL/6 b 2 2

FVB q 1 1

SJL s 2 1

F1(B6x129) b 2 2

F1 (B6xBALB/c) b/d 2 2

F1 (B6xCBA) b/k 2 2

B10;B6-Rag2-/-Il2rg-/- b 2 2

Table 2. Mouse strain characteristics

Research Strategy Page 42

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
become available for transplant a 

matching batch of MP could be selected 

and be available at the time of organ 

transplant. If the MP have to be derived 

from the organ donor itself the logistics 

become more complicated. Bone marrow 

cells would have to be harvested with the 

other organs from the donor and used 

either immediately following the 

transplant, after enrichment of progenitor 

and stem cells, or a week or two later 

after expansion of the MP at a central 

facility. Our preliminary data clearly 

indicates that full mismatching (major and 

minor transplantation antigens) leads to 

rejection: Tolerance induction by MP is specific. However, matching may not need to be absolute. BALB/c mice 

reconstituted with 4,000 FVB HSC and 106

 C57BL/6 MPc can accept skin grafts from B10;B6 mice (4/4 

accepted) while rejecting an AKR skin graft (0/1 accepted). While the degree of mismatching in this particular 

experiment is not clearly defined [70] it does indicate that there may be some leeway in the amount of 

matching required. We propose to determine the degree of mismatching tolerated in MP induced tolerance 

(Fig. 5).

Milestone: Determine the ability of standard doses of MP (106

 cells obtained after expansion) to protect grafts 

that are either fully matched to the MP (same inbred strain), matched in major MHC only (F1 to parent or HLAmatched strains) or are haploidentical to the MP. As acceptance rates may vary, and will not necessarily be 

100%, we will test fifteen mice per group (see data analysis section). If we fail to observe accepted grafts in 

mice receiving MP (ten or more rejected grafts) we will repeat that group using trachea grafts (if successful in 

aim 1) and an increased dose (5x106

 per mouse) of expanded MP cells. The group size will again be fifteen 

mice to give sufficient power (as detailed in the Data Analysis Section below).

Division of the work. The academic partner will inject MP and place and monitor skin grafts. Sorting of HSC, 

ex vivo expansion of HSC into MPc and cryopreservation of the MPc will be performed by the corporate 

partner. All cryopreserved MPc will undergo lot release testing that includes viability, composition, function, 

endotoxin levels, and bioburden to ensure a product with consistent purity and potency.

Model Host HSC donor MP donor Skin Graft 1 Skin Graft 2

Fully matched BALB/c FVB C57BL/6 C57BL/6 AKR

Majors BALB/c FVB AKR CBA AKR

Majors BALB/c FVB F1(B6x129) C57BL/6 AKR or 129

Haplo BALB/c FVB F1(B6xBALB/c) F1(B6xCBA) SJL or 129

Table 3. Strain distribution for Aim 2 experiments. Skin graft 1 and 2 represent the two skin grafts that can be placed simultaneously. 

They are accepted or rejected independently (submitted). Actual position (front or backward) will be assigned randomly. All strains 

(including F1’s) are directly available from approved vendors. To confirm that the results apply to lung transplants key experiments 

will be repeated using trachea grafts. Skin grafts are used initially because it is the most stringent mouse transplant model.

Details of the model. HSC and MP sorting and expansion will be as detailed under Aim 1, except that the 

mouse strains will be different (Table 3). On the day of injection, skin grafts will be placed that either are MPmatched, have matched major MHC antigens to the MP donor, are haploidentical to the MP-donor, or are fully 

mismatched. Skin grafts will be fully mismatched to host and HSC donors. The graft will either be a single skin 

graft (tail to belly) using our published protocol [3] or a double skin graft (two tail skin grafts to belly). Placement 

of the skin grafts on the same day as the HSC administration allows us to match the skin donor to the MP 

donor or HSC donor, which can be important when using mutant strains that may lack a fully inbred 

background. The fate of the tail skin will be monitored with digital photography, holding the non-anesthetized 

mouse at a fixed position over an upward facing camera with macro lens. In addition to skin grafts we will also 

test the model using trachea-grafts as detailed under Aim 1. Hematopoietic cells originating from host, HSC

donor and MP donor will be identified with 6-color flow cytometry using MHC class I or congenic markers 

(CD45) to determine origin.

Fig. 5. Experimental design for aim 2. BALB/c mice will be lethally 

irradiated, injected with 4,000 FVB HSC and 500,000 MPc (table 2), after 

which skin grafts or trachea grafts are placed (all on same day) from donors 

matched in varying degrees to the MP (table2). Primary outcome is 

skin/trachea graft acceptance or rejection. 

Research Strategy Page 43

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Potential Problems/Alternative Approaches. We may find that the level of tolerance achieved (defined as 

skin grafts not rejected) may not only depend on the degree of mismatching, but also on the number of MPs 

used. While that would not be a primary milestone, this may provide an opportunity to define the effects of 

increasing MP cell dose. In addition we are prepared to change MP culture conditions or to resort to using 

freshly purified MP from the bone marrow if we find it difficult to achieve tolerance with other than skin donorderived MP. We may also test the ability of MP using less easily rejected transplant models such as ear-heart 

transplantation, in which neonatal heart is transplanted into the pinna of the hosts ear [21]. While most 

experiments will be performed using MP from wild-type mice, some of the crucial data will be confirmed where

appropriate using MP from B10;B6-Rag2-/-

Il2rg-/- mice to confirm that only myeloid cells are essential.

Data Analysis. The primary variable in Aim 1 is epithelial maintenance/absence of occlusion based on 

histology and for Aim 2 skin graft acceptance or rejection based on gross morphology, confirmed by histology. 

Secondary variables are the number, origin, and types of cells present in reconstituted mice at various times, 

which will be expressed as absolute cell numbers (typical) or percent of cells and loss of epithelium in tracheas 

(expressed as percent of circumference without epithelium). Where necessary for parametric assumptions, 

appropriate transformations like log data will be employed and justified. If necessary, nonparametric tests such 

as the Wilcoxon rank sum test will be used. We will compare grafts using a paired t-test. Power: With 15 in 

each group at an alpha of 0.05 we will have at least 80% power to detect a difference in percentage of animals 

with the primary outcome of 0.6, assuming the grafts have a correlation of 0.2 or less. Although every effort will 

be made to avoid missing data, it is possible that some measurements will be unavailable. Whenever data are 

missing, the causes will be examined. Mice that die before analysis due to hematopoietic failure (<30 days 

post reconstitution) or infection (death <10 days post reconstitution) will be censored, as these experiments are 

not designed to study HCT-associated morbidity. In our experience MP administration does not increase 

mortality, and clinical studies are ongoing with MP to address this. All possible data will be included in the 

analysis by using appropriate methods to deal with missing data. Multiple imputations for missing items will be 

explored if the data are missing at random. These assays typically result in either clear rejection or acceptance 

of hypotheses for a given set of conditions. We will therefore take advantage of a two-step approach. Data will 

be analyzed using SPSS, Cytel StatXact, LogXact, and SAS version 9.01. For survival data we will use 

LogXact and Kaplan-Meier curves with a Wilcoxon logrank test

Timeline. Initial efforts will focus on 

Aim 1, starting with ex vivo expansion 

of MP. As these assays will be 

relatively rapid (with a read out 

typically at one month, though some 

of the mice may be followed longer) 

we expect to be able to analyze at 

least four iterations in six months, 

enabling us to meet the milestones 

defined for Aim 1. The assays for Aim 2 will take approx. two months to obtain results, and if more than a few 

iterations are needed will become be main focus of the second half of this Phase I project. We expect to meet 

the milestones described for Aim 2 9 to 12 months after initiation of the project.

Goals for Phase II. Once the milestones for Phase I have been met we plan to submit a proposal for Phase II 

funding with the focus in Phase II on two major aims. (1). The first aim would be to test human, clinical grade, 

MP for their ability to induce tolerance for human skin grafts in an immunodeficient mouse model. Specifically, 

we aim to test whether graft matched clinical grade human MP can prevent rejection of human skin 

transplanted to immunodeficient mice, by third party peripheral blood lymphocytes. (2) The second area of 

focus would be to define reduced intensity preconditioning regimens that allow induction of transplantation 

tolerance with MPc. The fact that MP induce tolerance for matched skin grafts when given with autologous, 

rather than allogeneic HSC indicates that sub-lethal preconditioning, with targeted destruction of host immune 

cells will be feasible. The information obtained in these aims, combined with the data from the proposed Phase 

I experiments, should allow us to design and implement trials to test this approach in patients. Clinical trials will 

be an obvious and essential step in commercializing MP for tolerance induction.

Fig 6. Proposed timeline for Phase I covering a 12 month period.

Research Strategy Page 44

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
VERTEBRATE ANIMALS 

1) For Cellerant Therapeutics (Laboratory of Dr. Karsunky)

Cellerant’s Animal Assurance Number: 

IACUC approved protocols for all animal work and procedures proposed under this proposal are already 

approved and in place at Cellerant. Prior to award Cellerant will amend those protocols to include the strains 

and number of animals needed for the work proposed here.

1. Description of proposed use 

This protocol involves the use of mice in order to study the therapeutic potential of human myeloid progenitor

cells (MPC) for the induction of operational tolerance to enable stable long term solid organ engraftment. We 

will use C57Bl/6, Balb/c, AKR, FVB, F1 (B/6x129), F1 (B/6xBalb/c) and B10;B6-Rag2-/-IL2rg-/- to isolate 

hematopoietic stem cells as the starting material for MPC ex vivo expansion cultures. All mice will be males of 

6-10 weeks of age at tissue harvest. 

Strain Number of animals estimated

C57Bl/6 100

Balb/c 100

AKR 100

FVB 100

F1 (B/6x129) 50

F1 (B/6xBalb/c) 50

B10;B6-Rag2-/-IL2rg-/- 40

Total 540

2. Justify the use of animal, choice of species and numbers used

For the isolation of hematopoietic stem cells (HSC) we depend on Thy1.1 expressing mouse strains. This is a 

less common allele but it is expressed in certain Bl/6 substrains, AKR and FVB mice. Therefore, these three 

were chosen as the main strains for HSC isolation as the starting material for MKP cultures. One of the key 

scientific questions will be to test for immunogeneic barriers for MPC to engraft and induce tolerance. For 

experiments proposed under Aim 1 we will use ex vivo derived MPC generated at Cellerant from FVB, C57Bl/6 

and AKR mice which are either syngeneic (matched) to the organ graft, matched to the HSC or unmatched to

both graft sources. To the actual degree of matching that is required between MPC and the graft we will 

generate MPC from C57Bl/6, AKR, F1 (B/6x129), and F1 (B/6xBalb/c) mice. B10;B6-Rag2-/-IL2rg-/- mice will 

be used to confirm that the tolerance induction seen from ex vivo expanded MPC is truly due to myeloid cells 

only without any contribution from lymphoid cells. We expect that the majority of our proposed experiments will 

be done using C57Bl/6, Balb/c, AKR, and FVB mouse strains, which is reflected in the higher mouse numbers 

for these strains.

The species mouse (Mus musculus) is clearly the best and probably only laboratory species where 

immunogenic barriers can readily be experimentally addressed. Mice are also a well established system for in 

vivo studies of the potential of hematopoietic stem and progenitor cells. The use of a standard laboratory 

species such as the mouse also helps to ensure optimal care of the animals because of the availability of wellestablished facilities and extensive veterinary expertise. The number of mice required reflects the need to 

generate and maintain the mice and to generate sufficient MPC for the planned engraftment studies. 

Experience with these models has shown that this is the number of mice necessary for study of each 

experimental parameter in order to convincingly answer the questions posed. 

3. Veterinary care

Cellerant Therapeutics has its own animal care facility. The facility complies with all applicable provisions of the 

Public Health Service Policy on Humane Care and Use of Laboratory Animals. The animal care staff are 

trained laboratory animal technologists and the investigators working on the experiments proposed in this 

application receive regular training on the proper handling and treatment of mice. 

Vertebrate Animals Page 45

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
4. Procedures for ensuring that discomfort is minimized

Animals used for the harvesting of bone marrow will be between 6-10 weeks of age. Mice will be housed as 

per Cellerant’s standard procedures. The mice will be euthanized through CO2 asphyxiation followed by

cervical dislocation. The femoral, coxae and tibiae bones will be removed and used as a source of bone 

marrow cells for HSC sorting. The carcasses will be disposed of as pathological material by approved methods 

of disposal. 

5. Methods of euthanasia

Mice will be sacrificed to obtain tissues e.g. for tissue analysis by flow cytometry and other assays. Mice will 

usually be euthanized by CO2 asphyxiation. In some instances mice may be anesthetized by isoflurane and the 

depth of anesthesia assessed by the withdrawal reflex, followed by cervical dislocation. If the reflex persists, 

additional anesthetic will be administered. 

2) For Children’s Mercy Hospital/University of Missouri, Kansas City (Laboratory of Dr. Domen)

1). Description of proposed use: This protocol involves the use of mice in order to study the tolerance 

induction/maintenance abilities of hematopoietic stem and progenitor cells. The strains used will include wildtype mice such as BALB/c, AKR, FVB, C57BL/6 (purchased from JAX) as well as mutant mice such as 

B10;B6-Rag2-/-

Il2rg-/- mice (purchased from Taconics) and other strains as listed and available from JAX. The 

ages will range from 5 weeks to 1-year-old adult mice. These mice will be used as sources for the purification 

of cells (HSC and MP) and as a source of tissue for skin transplants. In addition, mice will be used as 

recipients of both cells and skin transplants. All these experiments are aimed at developing conditions for 

inducing tolerance for skin transplantations.

2). Justify the use of animal, choice of species and numbers used: The experiments proposed in this 

proposal addresses the complex interactions between many different types of effector cells and molecules that 

occurs after transplantation of a non-matched solid organ. There are no protocols to mimic this in culture. This 

makes it impossible to predict the outcome of transplantations based on in vitro assays, despite the 

tremendous clinical value that such an assay would have. The mouse is the best model for the proposed 

studies. There are multiple well-characterized mouse strains available, wild-type and with defined mutations. 

Their hematopoietic stem and progenitor cells have been well characterized and can be sorted to near 

homogeneity using commercially available antibodies. Both congenic (same genetic makeup, typically except 

for a marker gene) and allogeneic (genetically different) models are available that allow the tracking of host 

and donor cells. In addition their response to radiation as a preconditioning regimen has been well 

documented, reducing the number of animals needed to set up the experiments. Solid organ transplant 

models, including ear-heart transplantation and skin graft transplantations, have been established. Most 

importantly, the results obtained in mice have historically translated well to clinical practice. No lower animal 

model has all of these features.

The primary variable in aim 1 is epithelial maintenance/absence of occlusion based on histology and for 

aim 2 skin graft acceptance or rejection based on gross morphology, confirmed by histology. Secondary 

variables are the number, origin, and types of cells present in reconstituted mice at various times, which will be 

expressed as absolute cell numbers (typical) or percent of cells and loss of epithelium in tracheas (expressed 

as percent of circumference without epithelium). Where necessary for parametric assumptions, appropriate 

transformations like log data will be employed and justified. If necessary, nonparametric tests such as the 

Wilcoxon rank sum test will be used. We will compare grafts using a paired t-test . Power: With 15 in each 

group at an alpha of 0.05 we will have at least 80% power to detect a difference in percentage of animals with 

the primary outcome of .6, assuming the grafts have a correlation of .2 or less. Ultimately we are interested in 

pair wise comparisons. We will not adjust for multiple comparisons. However, we will report the actual P values 

and repeat significant experiments for validation. 

With each group we expect to use approx 50 mice (15 hosts, 5 HSC donors, 30 trachea donors). We 

expect to use approx. 700 mice, 6 groups for aim 1, and 8 groups for aim 2. Transplanted mice will be followed 

for long periods of time (2 to 6 months) and will be analyzed (eg by flow cytometry of blood) regularly.

Vertebrate Animals Page 46

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
3). Veterinary care: The animal care facility of the University of Missouri Kansas City Medical School is 

fully accredited by the American Association for Accreditation of Laboratory Animal Care (AAALAC) and 

complies with all applicable provisions of the Public Health Service Policy on Humane Care and Use of 

Laboratory Animals. The animal care facility employs a veterinarian. The investigators working on the 

experiments proposed in this application receive regular training on the proper handling and treatment of mice. 

Animals which will be irradiated will be kept on antibiotic water prior to and after irradiation. 

Animal welfare assurance number: The techniques in this proposal have been IACUC 

reviewed and are currently approved as protocol 1112-Domen. Last approval date 11-01-2012.

4). Procedures for ensuring that discomfort is minimized: Total body irradiation used in the 

competitive repopulation assays will be performed in standard allentown cages in a RadSource 2000 X-ray 

machine. The mice will be exposed to a irradiation dose of 8-10 Gy total, depending on the mouse strain. All 

animals will be monitored for signs of distress (hunching, failure to groom, weight loss) and euthanized when 

moribund. Skin Transplantation Procedure. The surgeon's hands will be washed with hibiclens scrub 

(chlorhexidine) prior to putting on sterile gloves. Instruments will be prepared by immersion in clidox or 

sporocidin according to the manufacturers recommendations. A mouse will receive preoperative 

anesthetic/sedation by IP injection of xylazine/ketamine at 10/100mg/kg. The abdomen is prepped in a sterile 

fashion by shaving the hair using an Oster Animal Clipper. The mouse is then placed supine an on operating 

board and secured followed by prepping several times with hibiclens scrub. Eyes will be protected using 

antibiotic eye ointment. The skin is patted dry with sterile gauze prior to the incision. Sterile gauze or a sterile 

paper drape will be used to isolate the surgical field. A 1.5 X 1.5 cm segment of skin is removed, the 

subcutaneous tissues being left in place. Hemostasis is assured. The skin graft is then placed over the 

prepared recipient site, the size discrepancy between donor site and recipient site allowing the graft to be 

slightly stretched. Mouse grafts are placed such that the hair will grow in a direction incongruous with the 

surrounding hair. The skin is first secured at each of the four corners with 6-0 Maxon or Biosyn sutures. A 6-0 

Maxon or Biosyn is then used in an interrupted fashion to secure the graft to the surrounding skin. All fluid is 

decompressed from the graft site. A 2 X 2 piece of gauze is then sewn over the graft at its four corners with 4-0 

Sofsilk. The procedure is expected to last less then 20 min. The animal is closely observed until determined to 

be recovered from anesthesia. The respiratory rate and behavior of the animal will be monitored until the 

animal is fully recovered from anesthesia, moving around comfortably, and is able to drink easily. If recovery is 

prolonged a heat lamp, placed at least 15 inches away from the animals, will be used to prevent hypothermia. 

Buprenorphine is administered by s.c. injection at a dose of 0.05mg/kg twice daily starting immediately 

following recovery from anesthesia. Three injections are given routinely, more if the need is indicated by the 

animal. Limited discomfort is expected with collection of bloodsamples through nicking the tailvein. The 

limited discomfort does not justify the additional stress and risks associated with anesthesia. Trachea 

Transplant Model. Tracheas will be removed from transplant donors and kept in sterile ice-cold HBSS or PBS. 

A small piece on the back of recipient mice is shaved, prepped sterile and a subcutaneous pocket is 

generated. The trachea is placed and the skin closed with sutures. Up to four tracheas can be placed 

simultaneously on the same donor. The mice are euthanized, and the transplanted trachea removed for 

histological analysis, at different timepoints ranging from 7 days to 3 months post transplant. Implants in 

syngeneic mice or tolerant mice will retain their epithelial lining, while allogeneic transplants in animals that 

have not been tolerized will loose this. The airway will become fibrotic and close. This process is usually 

completed by 30 days, but may take longer in the context of hematopoietic reconstitution.

5). Methods of euthanasia: Mice will be sacrificed to obtain tissues for staining, FACS sorting, to 

harvest skingrafts and for terminal analysis. Consistent with the guidelines, the mice will first either be 

anesthetized by intraperitoneal injection using ketamine (200mg/kg) plus xylazine (10mg/kg) or by CO2

asphyxiation. The depth of anesthesia will be assessed by the withdrawal reflex. If the reflex persists, 

additional anesthetic will be administered. All mice then undergo cervical dislocation. The mouse will be 

exsanguinated by cardiac puncture only if peripheral blood is wanted.

Vertebrate Animals Page 47

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
Select Agent Research 

Not applicable.

Select Agent Research Page 48

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
MULTIPLE PD/PI LEADERSHIP PLAN 

Rationale. This STTR application has two project sites. The small business project site is at 

Cellerant Therapeutics in San Carlos, CA. Holger Karsunky, PhD, will be the Principal 

Investigator at Cellerant Therapeutics. The academic project site is at Children’s Mercy Hospital 

and Clinics/University of Missouri Kansas City in Kansas City, MO, and Jos Domen PhD will be 

the on-site Principal Investigator and direct the efforts 

Communication Plan. The two project leaders have talked and met regularly over the last 7 

years. In the context of this project these contacts would be intensified and scheduled on a 

regular basis to include a standing tele conference at least every two weeks to discuss the 

progress towards the project’s milestones. It is also planned to have reciprocal visits to further 

strengthen the collaboration and hold one face-to-face kick-off meeting at the beginning of the 

performance period as well as one in the middle of the project to discuss progress and the 

strategy to apply for continued funding. 

Role. Dr. Karsunky will lead in coordinating the efforts related to this project at Cellerant 

Therapeutics, and in determining how this project will fit into the commercial development of 

Cellerant Therapeutics. Dr. Domen will lead the efforts at Children’s Mercy Hospital and 

Clinics/University of Missouri Kansas City, and is responsible for the in vivo preclinical tests that 

are necessary to determine the parameters under which this novel indication for myeloid 

progenitor use can be tested and used clinically. 

Conflict Resolution. Conflicts will be resolved in consultation between the two PI’s. Decisions 

on the commercial use and development of this technology, the eventual aim of this project, by 

Cellerant Therapeutics will rest with Dr. Ram Mandalam the CEO of Cellerant Therapeutics. 

According to SF424 guidelines the small business concern will be the primary party that will 

exercise management direction and control of the performance of the project. 

Budget allocation. Half of the budget is currently allocated to each of the project sites, 

conforming to the requirement that at least 40% of the work is performed at the business site 

and at east 30% at the academic site. This will allow both groups to contribute significantly in 

this first phase of the transfer of this technology from the Domen Lab to Cellerant Therapeutics.

Multiple PI Leadership Plan Page 49

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
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RESOURCE/DATA SHARING PLAN 

Data Sharing Plan 

Based on the SF424 (R&R) SBIR/STTR Application Guide for NIH and Other PHS Agencies a formal Data 

Sharing does not need to be provided since the requested direct costs are under $ Data generated by 

this project will be shared via publication in peer reviewed journals. Additionally, study team members may 

present the findings in lectures or posters at conferences and seminars. 

Sharing Model Organisms 

The proposed work does not include the development of new model organisms. Only mouse strains that are 

readily available from commercial sources will be used for the proposed work. 

Policy for Genome-Wide Association Studies 

No genome-wide association studies (GWAS) to identify common genetic factors are planned as part of this 

proposal. 

Resource Sharing Plan Page 57

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
PHS 398 Checklist

OMB Number: 0925-0001 

1. Application Type:

From SF 424 (R&R) Cover Page. The responses provided on the R&R cover page are repeated here for your reference, as you answer 

the questions that are specific to the PHS398.

* Type of Application:

Federal Identifier: 

2. Change of Investigator / Change of Institution Questions

Change of principal investigator / program director

Name of former principal investigator / program director: 

Change of Grantee Institution

* Name of former institution:

3. Inventions and Patents (For renewal applications only)

* Inventions and Patents:

If the answer is "Yes" then please answer the following:

* Previously Reported: 

Prefix:

* First Name:

Middle Name:

* Last Name:

Suffix:

New Resubmission Renewal Continuation Revision

Yes No

Yes No

	


 		

 ! " 

Checklist Page 58

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger
4. * Program Income

If you checked "yes" above (indicating that program income is anticipated), then use the format below to reflect the amount and 

source(s). Otherwise, leave this section blank.

Is program income anticipated during the periods for which the grant support is requested?

*Budget Period *Anticipated Amount ($) *Source(s)

Yes No

5. * Disclosure Permission Statement

If this application does not result in an award, is the Government permitted to disclose the title of your proposed project, and the name, 

address, telephone number and e-mail address of the official signing for the applicant organization, to organizations that may be 

interested in contacting you for further information (e.g., possible collaborations, investment)? 

Yes No

	


 		

 ! " 

Checklist Page 59

Principal Investigator/Program Director (Last, first, middle): Karsunky, Holger