SUNY Downstate Health Sciences University
Department of Pathology
Matthew R. Pincus, PhD
Professor of Pathology
Chief, Department of Pathology & Laboratory Medicine
New York Harbor VA Medical Center
Tel: (718) 270-3688 • Fax: (718) 270-2960
Department of Pathology
SUNY Downstate Medical Center
Department of Pathology & Laboratory Medicine
New York Harbor VA Medical Center,
800 Poly Place
Brooklyn, NY, 11203
Research Interest Summary
We have designed a group of anti-cancer peptides from the ras-p21 and p53 proteins that kill cancer cells but have no effect on normal cells.
Our inventions encompass 2 sets of peptides, one from the p53 anti-onogene protein and one from ras-p21 oncogenic protein. We have designed two sets of peptides from the ras-p21 and p53 proteins using computer-based molecular modeling of the three-dimensional structures of these proteins. We have synthesized three p53 peptides from its mdm-2-binding domain (i.e., residues 12-26, 17-26 and 12-20) each attached to a pentratin sequence that allows for its transport across the cell membrane. P53 12-26-penetratin, p53 17-26-penetratin and p53 12-20-penetratin are called PNC-27, 28 and 21, respectively. We first tested these peptides against two cell lines that we developed: a normal pancreatic acinar cell line, called BMRPA1, and its malignantly transformed counterpart cell line, called TUC-3, which we developed by transfecting the ras-oncogene into these BMRPA1 cells. When we incubated any of these peptides with TUC-3 cells, we found that all of them, but not a control peptide called PNC-29, killed all of the TUC-3 cells within 3 days but had no effect on the growth or viability of BMRPA1 cells. Significantly, we found that PNC-28 (p53 17-26-penetratin) had no effect on the ability of human stem cells from cord blood to differentiate into hematopoietic cell lines suggesting that this peptide would not suppress bone marrow. We have tested PNC-27 and 28 on 20 different human cancer cell lines and found that they are cytotoxic to all of them, inducing total cancer cell death in very short periods of time. For example, PNC-27 kills three different human breast cancer cell lines within 1 hour while having no effect on an untransformed human breast epithelial cell line. We have obtained evidence that the peptides work by interacting with two proteins that are present in the cell membranes of cancer but not normal cells. The peptides then induce membranolysis of the cancer cells. We have further tested our peptides against the highly malignant, metastatic TUC-3 pancreatic cancer cell line in nude mice and find that they eradicate the tumor within two weeks of drug delivery. This work has been published as the cover article in the September, 2006 issue of the International Journal of Cancer. It appears that this peptide would prove successful in treating a variety of human cancers.
We have further designed two peptides from the ras-p21 oncogenic protein using computer-based molecular modeling. These peptides correspond to amino acid residues 35-47 (PNC-7) and 96-110 (PNC-2). These two peptides selectively block oncogenic ras-p21 but not wild-type ras-p21 in a Xenopus (frog) oocyte model system. We have found that wild-type and oncogenic ras-p21 utilize different signal transduction pathways allowing for the oncogenic pathway to be selectively inhibited. We then proceeded to test these peptides on a ras-transformed pancreatic cancer cell line. In these experiments both peptides were linked to a penetratin peptide sequence that allows the peptide to be transferred across the cell membrane. This cell line, called TUC-3, was produced by transfecting the ras oncogene into a normal pancreatic acinar cell line, called BMRPA1. When we incubated either peptide with TUC-3 cells, the cells reverted back to the untransformed phenotype, i.e., became normal cells again. On the other hand, treatment of the normal acinar cells with either peptide had no effect either on their growth rates or on their viabilities. We have tested these two peptides on a number of different human cancer cell lines, i.e., fibrosarcoma, colon cancer, pancreatic cancer and astrocytoma, and found that both peptides either induce reversion to the untransformed phenotype or induce cancer cell death.
Funded by: VA and the American College of Surgical Oncology
- Michl, J., Scharf, B., Schmidt, A., Hannan, R., von Gizycki, H., Friedman, F.K., Brandt-Rauf, P.W., Fine, R.L. and Pincus, M.R. (2006) PNC-28, a p53 Peptide that Is Cytotoxic To Cancer Cells, Blocks Pancreatic Cancer Cell Growth in vivo. Int. J. Cancer, 119, 1577-1585.
- Qu, Y., Adler, V., Izatova, L., Pestka, S., Bowne, W., Michl, J., Boutjdir, M., Friedman, F.K. and Pincus, M.R. (2007) The dual-specificity kinases, TOPK and DYRK1A, are critical for oocyte maturation induced by wild-type-but not by oncogenic- ras-p21 protein. Frontiers in Bioscience 12, 5089-5097.
- Pincus, M.R., Michl, J, Bowne, W. and Zenilman, M. (2007) Anti-Cancer Peptides from the ras-p21 and p53 Proteins. Research Advances in Cancer. R.M. Mohan, Ed. Global Research Network Publishers, Kerala, India, 65-90.
- Bowne, W., Michl, J., Bluth, M.H., Zenilman, M. and Pincus, M.R. (2007) Novel Peptides from the RAS-p21 and p53 Proteins for the Treatment of Cancer. Cancer Ther. 5, 331-346.
- Monaco, R., Rosal, R., Dolan, M.A., Pincus, M.R. and Brandt-Rauf, P.W. (2008) Conformational Effects of a Common Codon 399 Polymorphism on the BRCT1Domain of the XRCC-1 Protein. Protein J., in press.
- Adler, V., Bowne, W., Kamran, I., Michl, J., Friedman, F.K., Chin, E., Zenilman, M. and Pincus, M.R. (2007) Two Peptides Derived from ras-p21 Induce Either Phenotypic Reversion or Tumor Cell Necrosis of ras-Transformed Human Cancer Cells. Cancer Chemother Pharmacol. 62, 491-498.
- Adler, V., Bowne, W., Michl, J., Ikram, K., Pestka, S., Izatova, L., Zenilman, M., Friedman, F.K. and Pincus, M.R. (2007) Site-Specific Phosphorylation of raf in ras-Transformed Cells Mediated by jun-N-Terminal Kinase. Ann Clin Lab Med 38, 47-56
- HooKim, K., deRoux, S., Igbokwe, A., Stanek, A., Koo, J., Hsu, J., Pincus, M.R. and Bluth, M. H. (2007) IgG Anti-Cardiomyocyte Antibodies in Giant Cell Myocarditis. Ann clin Lab Med 38, 83-87.
- Cai, C.Q., Peng, Y., Buckley, M.T., Wei, J., Chen, F., Liebes, L., Gerald, W.L., Pincus, M.R., Osman, I. and Lee, P. (2007) Epidermal Growth Factor Receptor Activation in Prostate Cancer by Three Novel Missense Mutations. Onocogene 27, 3201-3210.