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DCGT research priorities:Led by Critical Path Challenges

VirologyRetroviruses, lentivirus, adeno, filovirus

ImmunologyAnti-viral immunity, immunobiology of cell therapy and

xenotransplantationCell biology

Control of differentiation in animal models, stem cell biology

Cancer biologyAntigen characterization, Molecular biomarkers, animal models

Biotechnology - Genomics, flow cytometry, proteomicsTissue Safety Program

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Division of Cellular and Gene TherapiesRaj Puri, Ph.D., M.D., Division Director

Kimberly Benton, Ph.D., Deputy Director

Division of Cellular and Gene TherapiesRaj Puri, Ph.D., M.D., Division DirectorKimberly Benton, Ph.D., Deputy Director

Cellular and Tissue Therapy Branch

Steven Bauer, Ph.D., Chief

Tumor Vaccines and Biotechnology Branch

Raj Puri, Ph.D., M.D., Chief

Cell Therapies BranchKeith Wonnacott, Ph.D., Chief

Gene Therapies BranchDaniel Takefman, Ph.D., Chief

Gene Transfer and Immunogenicity Branch

Andrew Byrnes, Ph.D., Chief

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Tumor Vaccines andBiotechnology Branch (TVBB)

Raj K. Puri, M.D., Ph.D. Cancer Biology and Genomics Program

Syed R. Husain, Ph.D., Staff ScientistBharat H. Joshi, Ph.D., Staff Scientist

Michail Alterman, Ph.D. Proteomics Program

Shyh-Ching Lo, M.D., Ph.D.Tissue safety Program

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Scientific and Regulatory Scientific and Regulatory Challenges in cancer Vaccines and Challenges in cancer Vaccines and

Immunotherapy Immunotherapy – Puri lab– Puri lab

Complex biology of cancer Complex biology of cancer Identification of appropriate targets and Identification of appropriate targets and

specificity of targetspecificity of target Appropriate tests and biomarkers for identity and Appropriate tests and biomarkers for identity and

potency of cancer therapeutics/vaccines potency of cancer therapeutics/vaccines Animal models – safety and efficacyAnimal models – safety and efficacy Immune biomarkers of response and disease Immune biomarkers of response and disease

monitoringmonitoring

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Research programs addressingResearch programs addressingscientific and regulatory challengesscientific and regulatory challenges

Specific Programs:Specific Programs: Characterization of tumor associated cell surface Characterization of tumor associated cell surface

proteins (antigens or receptors) for identity and potency proteins (antigens or receptors) for identity and potency and target these receptors for cancer therapyand target these receptors for cancer therapy

Animal models of human cancer to assess safety and Animal models of human cancer to assess safety and efficacy of tumor targeted agents including cancer efficacy of tumor targeted agents including cancer vaccines, immunotoxins, cell and gene therapy productsvaccines, immunotoxins, cell and gene therapy products

Characterization of cancer vaccines and embryonic stem Characterization of cancer vaccines and embryonic stem cells by genomics technology to identify cancer stem cells by genomics technology to identify cancer stem cells and biomarkers of identity and potency and cells and biomarkers of identity and potency and response to cancer vaccinesresponse to cancer vaccines

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Characterization of tumor associated cell Characterization of tumor associated cell surface receptors and antigenssurface receptors and antigens

Discovery of overexpression of Th-2 derived cytokine Discovery of overexpression of Th-2 derived cytokine receptors in tumorsreceptors in tumors

Interleukin-4 receptorsInterleukin-4 receptors RCC, malignant glioma, AIDS-Kaposi’s sarcoma, colon RCC, malignant glioma, AIDS-Kaposi’s sarcoma, colon

cancer, breast carcinoma, NSCLC, prostate cancer, ovarian cancer, breast carcinoma, NSCLC, prostate cancer, ovarian carcinoma, mesothelioma, hematological malignancies carcinoma, mesothelioma, hematological malignancies (CLL-B),(CLL-B), PDA, biliary ductal cancer and SCCHNPDA, biliary ductal cancer and SCCHN

Interleukin-13 receptorsInterleukin-13 receptors RCC, malignant glioma, pediatric glioma, AIDS-Kaposi’s RCC, malignant glioma, pediatric glioma, AIDS-Kaposi’s

sarcoma, SCCHN, ovarian carcinoma, PDA, sarcoma, SCCHN, ovarian carcinoma, PDA, pheochromocytoma, brain stem glioma and prostate cancerpheochromocytoma, brain stem glioma and prostate cancer

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Animal models of human cancer to assesssafety, toxicity and efficacy of cancer

targeted agents

Ovarian Cancer Pancreatic cancer Glioblastoma multiforme SCCHN Mesothelioma Lung cancer

AIDS-Kaposi’s tumors Breast cancer Hodgkin’s lymphoma Melanoma Pheochromocytoma

Pheochromocytoma

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Conclusions and Ongoing ProjectsConclusions and Ongoing Projects Certain human tumors express high levels of IL-4R and IL-13R Certain human tumors express high levels of IL-4R and IL-13R in in

vitrovitro and and in vivo; in vivo; the structure and signal transduction through these the structure and signal transduction through these receptors is different in cancer and normal cells.receptors is different in cancer and normal cells.

IL-4R and IL-13R can be effectively targeted by immunotoxins by IL-4R and IL-13R can be effectively targeted by immunotoxins by themselves or when combined with other agents for cancer therapy. themselves or when combined with other agents for cancer therapy.

IL-13RIL-13R2 is involved in PDA invasion and metastasis and may be a 2 is involved in PDA invasion and metastasis and may be a prognostic biomarker of disease. prognostic biomarker of disease.

IL-13RIL-13R2 can signal through AP-1 pathway and regulated by 2 can signal through AP-1 pathway and regulated by Adrenomedullin Adrenomedullin in vitroin vitro and and in vivo.in vivo.

IL-13RIL-13R2 is a novel tumor rejection antigen - IL-13R2 is a novel tumor rejection antigen - IL-13R2 cDNA 2 cDNA vaccination induces immune response in preventive and therapeutic vaccination induces immune response in preventive and therapeutic murine cancer models. murine cancer models.

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Analytical proteomics for the characterization of biologic products and discovery of

biomarkers

Michail Alterman, Ph.D.Michail Alterman, Ph.D.Senior Investigator, Tumor Vaccines and Biotechnology Branch,

Division of Cellular and Gene Therapies

Laboratory Goal:Laboratory Goal: Development and application of mass spectrometry-Development and application of mass spectrometry-

based proteomic tools for qualitative and based proteomic tools for qualitative and quantitative measurement of cellular products, cell quantitative measurement of cellular products, cell substrates and vaccines and discovery of potential substrates and vaccines and discovery of potential biomarkers of cancerbiomarkers of cancer

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Mission relevant questions to be impacted by application of proteomics tools

Implementation of new better, clearer and more defined means for regulatory decision making in evaluation and review of cell substrates, vaccine products and other biologics

Development of valid protein biomarkers for examining drug safety and efficacy through prediction of therapeutic effect, selection of appropriate dose, and identification of risk of toxicity.

Development of improved strategies for biotechnology product physicochemical comparability assessments

Development of effective tools to be used in biologics and protein therapeutics manufacturing process controls and release testing

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Specific aimsSpecific aims and achievements since Site Visit in April 2010and achievements since Site Visit in April 2010

Specific Aim # 1. Development of mass spectrometry-based analytical tools for testing of biological product quality and identity.

Manuscript published in Analytical Biochemistry

Specific Aim # 2. Identification of proteomics-based cellular molecular signatures that may be tested as predictors of therapeutic success

and be developed as biomarkers of safety and effectiveness in preclinical animal models.

Invited talk at an international meeting, one poster presented at an international meeting and one at a national meeting, two manuscripts

in preparation

Specific Aim #3.Specific Aim #3. Proteomics-based analysis of influenza virus and Proteomics-based analysis of influenza virus and vaccines.vaccines.

One invited seminar, manuscript in preparationOne invited seminar, manuscript in preparation

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ConclusionsConclusions

A new label free quantitation technique was developed A new label free quantitation technique was developed that allows determination of a ratio of co-migrating in gel that allows determination of a ratio of co-migrating in gel electrophoresis proteinselectrophoresis proteins

Two prospective protein groups were identified that could Two prospective protein groups were identified that could be used as biomarkers for characterization of cell be used as biomarkers for characterization of cell substrates substrates

MS-based proteomics methods can be used to MS-based proteomics methods can be used to identify/distinguish different influenza variants and clades identify/distinguish different influenza variants and clades and quantify proteins in flu vaccines and virus and quantify proteins in flu vaccines and virus preparationspreparations

A comprehensive proteome map of human MSC is under A comprehensive proteome map of human MSC is under development; a number of prospective biomarkers of cell development; a number of prospective biomarkers of cell sub-culturing was found among >4000 identified proteinssub-culturing was found among >4000 identified proteins

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Ongoing projectsOngoing projects

1.1. Development of a new potency/stability Development of a new potency/stability assay for flu vaccine.assay for flu vaccine.

2.2. MSC project: MSC project:

i. identify protein signatures characteristic i. identify protein signatures characteristic for MSC from multiple donorsfor MSC from multiple donorsii. identify molecular signatures that could ii. identify molecular signatures that could be used to discriminate product that be used to discriminate product that produced a desirable clinical outcomeproduced a desirable clinical outcome

Tissue Microbiology Laboratory Tissue Microbiology Laboratory for Safety of Human Tissues for Safety of Human Tissues Intended for TransplantationIntended for Transplantation

Shyh-Ching Lo, Ph.D., M.D.Shyh-Ching Lo, Ph.D., M.D.

SV Report (2010) for The Tissue Laboratory Program of Tumor Vaccines and Biotechnology Branch

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Public Health, Regulatory Challenges and Public Health, Regulatory Challenges and Critical Path Scientific Research: Critical Path Scientific Research:

To Better Understand The Scientific Trends and Further To Better Understand The Scientific Trends and Further Strengthen The Dialogue with The Stakeholders of Human Strengthen The Dialogue with The Stakeholders of Human

TissueTissue Increasing number of tissues are recovered and processed as Increasing number of tissues are recovered and processed as

grafts by the industry and used by the medical practice each year. grafts by the industry and used by the medical practice each year.

The OCTGT within CBER regulates human cells, tissues, and The OCTGT within CBER regulates human cells, tissues, and cellular and tissue-based products (HCT/Ps) in order to prevent cellular and tissue-based products (HCT/Ps) in order to prevent the spread or transmission of communicable diseases. the spread or transmission of communicable diseases.

The scientific capability of the new program needs to cover a wide The scientific capability of the new program needs to cover a wide spectrum of expertise with very different microbial pathogens, spectrum of expertise with very different microbial pathogens, including bacteria, fungi, viruses and protozoa parasites.including bacteria, fungi, viruses and protozoa parasites.

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Research Programs of Tissue Research Programs of Tissue LaboratoryLaboratory

Specific Aims:Specific Aims: To establish the required scientific capabilities to directly To establish the required scientific capabilities to directly

support regulatory needs for tissue safetysupport regulatory needs for tissue safety

To adopt new technologies for rapid detection of infectious To adopt new technologies for rapid detection of infectious pathogens with high sensitivity in human tissues being pathogens with high sensitivity in human tissues being processed for transplantationprocessed for transplantation

To evaluate innovative methods for effective inactivation of To evaluate innovative methods for effective inactivation of various pathogens while preserving tissue quality for clinical various pathogens while preserving tissue quality for clinical needsneeds

To explore new scientific approaches for detection and To explore new scientific approaches for detection and characterization of previously unknown or newly emerging characterization of previously unknown or newly emerging infectious pathogens that would likely threaten the safety of infectious pathogens that would likely threaten the safety of human tissue graftshuman tissue grafts

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Laboratory research projects initiated Laboratory research projects initiated to accomplish the specific aims:to accomplish the specific aims:

Establishment of the laboratory capability of clinical microbiology Establishment of the laboratory capability of clinical microbiology for detection and characterization of microbes with tissue safety for detection and characterization of microbes with tissue safety concernsconcerns -- Investigation of incidents and validation of industrial -- Investigation of incidents and validation of industrial sterilization methodssterilization methods

Development of rapid, highly sensitive technology for detection of Development of rapid, highly sensitive technology for detection of pathogens in tissues intended for transplantation -- pathogens in tissues intended for transplantation -- RReal-time eal-time qPCR arrays of the targeted high-risk bacteria and RCDAD virusesqPCR arrays of the targeted high-risk bacteria and RCDAD viruses

Development of high-throughput genomic sequencing capability Development of high-throughput genomic sequencing capability for detection and characterization of previously unknown for detection and characterization of previously unknown infectious pathogens in tissue or tissue-based products infectious pathogens in tissue or tissue-based products -- -- Readiness of scientific capability against potential newly emerging Readiness of scientific capability against potential newly emerging threats of public healththreats of public health

Identification of “biomarkers” associated with the injury Identification of “biomarkers” associated with the injury mechanisms in cell/tissue and development of the injury-mechanisms in cell/tissue and development of the injury-associated microarray -- associated microarray -- Assessment of the degree of tissue injury Assessment of the degree of tissue injury in pathogen inactivation using various chemical treatmentsin pathogen inactivation using various chemical treatments

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Conclusions and Ongoing ProjectsConclusions and Ongoing Projects

A new tissue microbiology laboratory has been A new tissue microbiology laboratory has been established under the DCGT and the DHT to established under the DCGT and the DHT to enhance both the safety and the availability of enhance both the safety and the availability of high quality human tissue intended for high quality human tissue intended for transplantation.transplantation.

Development of New Technologies and Development of New Technologies and evaluation of innovative methods is being evaluation of innovative methods is being implemented to guard the safety and quality of implemented to guard the safety and quality of tissue intended for transplantationtissue intended for transplantation

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Thank you