Laboratory of Howard Kaufman, MD
Principal Investigator
Howard Kaufman, MD
- Residency, Boston University
- Surgical Oncology Fellowship, National Cancer Institute
- Tumor Immunology, National Cancer Institute
Background for Research focus:
My laboratory focuses on the development of recombinant poxviruses as vaccines for the treatment of cancer. Initial studies have documented that poxviruses engineered to express tumor antigens can be used to prime T cell responses in both murine models and in clinical trials. However, clinical responses are infrequent. One obstacle to effective tumor immunity is the suppression of T cell responses in the tumor microenvironment. We are now exploring the local delivery of recombinant poxvirus vaccines expressing costimulatory molecules or chemokines as a general strategy for manipulating local T cell immunity. This work involves molecular virology and poxvirus biology, a murine model of spontaneous colorectal cancer, and clinical trials. A component of the laboratory is dedicated to monitoring T cell responses in patients treated with a variety of vaccine and immunotherapy regimens. The monitoring includes in vitro cellular assays, real time PCR, microarray and proteomics analyses.
Main Focus of the Laboratory:
Development of recombinant poxviruses as tumor vaccines, use of co-stimulatory molecules and chemokines as vaccine adjuvants. Understanding mechanisms of tumor rejection in transgenic mice. Clinical application of tumor vaccines and monitoring anti-tumor immunity in cancer patients.
Please see our lab website at www.tumorvaccines.com.
Current Projects:
1. Clinical Vaccine Studies
A variety of clinical trials have been initiated using recombinant poxviruses as vaccines for patients with colorectal cancer, prostate cancer, renal cell carcinoma, pancreatic cancer, breast cancer, and malignant melanoma. The vaccine trials include an analysis of patient T cell responses before and after vaccination testing the hypothesis that vaccination induces antigen-specific immunity.
Immune assays include measurement of antibody titers, T cell precursor frequency by ELISPOT assay, and intracellular cytokine staining. We have recently embarked on a series of clinical trials to test the local delivery of costimulatory molecules into tumor cells by recombinant poxvirus vaccines. As part of these studies we have collected RNA from injected lesions and are using real time PCR and microarray analysis to identify specific immune genes that may be predictive of clinical responders. This will be correlated with proteomics profiling of banked serum from vaccinated patients.
2. Development of New Recombinant Vaccines
Several new recombinant vaccines have been developed in the lab. Vectors expressing CD40 Ligand and 4-1BBL have been engineered and are being tested in in vitro and murine tumor models. A major goal of this project is to understand the mechanism by which these vaccines exert anti-tumor properties in a murine model of colon cancer. These studies will help guide the design of clinical trials.
3. Chemokines
Our laboratory has engineered a number of recombinant poxviruses expressing chemokines. The lymphoid chemokine, CCL21 attracts naïve T cells and mature dendritic cells and mediates tumor regression when expressed by vaccinia virus. However, our lab now has evidence that CCL21 may act as a costimulatory molecule regulating the activation and differentiation of T cells. We are attempting to define exactly how CCL21 promotes T cell priming and influences the immune response.
4. Transgenic Mouse Model
A major issue in using CEA as a vaccine target is the possible role of pre-existing tolerance to CEA since it represents a self-antigen. Previous animal models used human CEA expressed in murine colon cancer cells and cannot be used to adequately evaluate the role of tolerance and immunity associated with CEA vaccines. A more appropriate animal model would be a transgenic mouse that exhibits a similar pattern of oncofetal human CEA expression and also develops malignant CEA-expressing tumors at a predictable rate. A mouse containing the human CEA transgene has been constructed which shares a similar spatio-temporal expression pattern with human CEA in an H-2b background.
Familial polyposis is a disorder characterized by the development of multiple adenomatous polyps throughout the colon of affected individuals and invariably leads to carcinoma by the age of 40. The disease has been linked to mutations of the adenomatous polyposis coli (APC) gene, which may also be responsible for some sporadic colon cancers. A transgenic mouse (H-2b) has been generated that contains a truncated segment of the murine APC gene with only 1638 base pairs present (APC 1638). These mice develop gastrointestinal tumors between 8 and 10 months of age and share a similar histologic appearance to human colon adenocarcinomas. We are generating a hybrid mouse expressing both the human CEA and murine APC 1638 transgenes would be ideal for the development of a model system wherein human CEA is expressed in the same spatio-temporal fashion as in human patients and where gastrointestinal tumor form on a regular, predictable basis. These mice will be characterized for transgene expression using PCR assays. Histologic evaluation will also be performed to determine the rate of tumor formation and progression as well as the induction of cell mediated immunity after vaccination. These mice will also be used for evaluating CEA-directed vaccine therapy and establishing the role of CEA tolerance in this model system.
Recent Publications:
Horig, H., Lee, D.S., Conkright, W., Park, D., Hasson, H., Rivera, A., LaMare, M., Tine, J., Guito, K., Tsang, K.-Y., Schlom, J., and Kaufman, H.L. B7.1 costimulation enhances the response of a recombinant canarypoxvirus (ALVAC) vaccine expressing human carcinoembryonic antigen (CEA). Cancer Immunol Immunother 49:504-514, 2000.
Kicska, G.A., Long, L., Horig, H., Fairchild, C., Tyler, P.C., Furneaux, R.H., Schramm, V.L., and Kaufman, H.L. A powerful transition state analog inhibitor of purine nucleoside phosphorylase selectively inhibits human T lymphocytes. Proc Natl Acad Sci USA 98:4593-4598, 2001.
Horig, H., Wainstein, A., Long, L., Kahn, D., Soni, S., Marcus, A., Edelmann, W., Kucherlapati, R., and Kaufman, H.L. A new mouse model for evaluating the immunotherapy of human colorectal cancer. Cancer Res 61:8520-8526, 2001.
Lieberman, S. Horig, H., and Kaufman, H.L. Innovative treatments for pancreatic cancer. Surg Clin N Amer 81:715-739, 2001.
Kaufman, H.L., DeRaffele, G., Divito, J., Horig, H., Lee, D., Panicali, D., and Voulo, M. A phase I trial of intra-lesional rV-TRICOM vaccine in the treatment of malignant melanoma. Human Gene Ther. 12:1459-1480, 2001.
Lee, H., Woodman, S.E., Engelman, J.A., Volonte, D., Galbiati, F., Kaufman, H.L., Lublin, D.M., and Lisanti, M.P. Palmitoylation of caveolin-1 at a single site (cys-156) controls its coupling to the c-src tyrosine kinase. J Biol Chem 276:35150-35158, 2001.
Eppler, E., Horig, H., Kaufman, H.L., Groscurth, P. and Filgueira, L. Antigen presentation and specific T-cell priming after transfection of human dendritic cells with mRNA encoding carcinoembryonic antigen. Eur J Cancer 38:184-193, 2002.
Kaufman, H.L., Swartout, B., Horig, H., and Lubensky, I. Combination interleukin-2 and interleukin-12 induces severe gastrointestinal toxicity and epithelial cell apoptosis in mice. Cytokine 17:43-52, 2002.
Kaufman, H.L., Flanagan, K., Lee, C.S.D., Perretta, D.J., and Horig, H. Insertion of interelukin-2 (IL-2) and interleukin-12 (IL-12) genes into vaccinia virus results in effective anti-tumor responses without toxicity. Vaccine 20:1862-1869, 2002.
Kaufman, H.L., Cheung, K., Haskall, Z., Horig, H., Hesdorffer, C., Panicali, D., DeRaffele, G., and Spanknebel, K. Intra-lesional rF-B7.1 versus rF-TRICOM vaccine in the treatment of metastatic cancer. Human Gene Ther 14:803-827, 2003.
Kaufman, H.L., Wei W., Manola, J., DiPaola, R.S., Ko, Y., Sweeney, C., Whiteside, T.L., Schlom, J., Wilding, G., and Weiner, L.M. Phase II Randomized Study of Vaccine Treatment of Advanced Prostate Cancer (E7897): A Trial of the Eastern Cooperative Oncology Group. J Clin Oncol 11:1-12, 2004.
Flanagan, K., Glover, R.T., Horig, H., and Kaufman, H.L. Local delivery of recombinant vaccinia virus expressing secondary lymphoid chemokine (SLC) results in a CD4+ T cell dependent antitumor response. Vaccine In press, 2004.
Kaufman, H. and Disis, N. Immune system versus tumor: Shifting the balance in favor of dendritic cells and effective immunity. J Clin Invest 113:664-667, 2004.
Bereta, J., Bereta, M., Park, J., Medina, F., Kwak, H., and Kaufman, H.L. Immune Properties of Recombinant Vaccinia Virus Encoding CD154 (CD40L) are Determined by Expression of Virally Encoded CD40L and the Presence of CD40L Protein in Viral Particles. Cancer Gene Ther In press, 2004.
Recent (last 5 years) Surgery Residents (Columbia and outside):
1. Giovanni Cessani, MD General Surgery Resident, University of Milan, Italy
2. John Stoutenburg, MD Medical Oncology Fellow, Columbia University
3. Ian Komenaka, MD Surgical Oncology Fellow, Columbia University
4.Alberto Wainstein, MD General Surgery, University of Belo Horizonte, Belo Horizonte, Brazil
5. Christopher Lee, MD Urology Resident, Albert Einstein College of Medicine
Back to Top