SUNY Downstate Health Sciences University
Department of Pathology
Alejandro Zuretti, MD
Department of Pathology
Tel: (718) 270-1643
Education and Training
- 1971-1976 National College of Buenos Aires, Buenos Aires, Argentina, Bachelor's Degree, Magna cum laude
- 1977-1983 School of Medicine, University of Buenos Aires, Argentina Doctor of Medicine, Cum Laude
- 1985-1986 Internship in Internal Medicine, St. John’s Episcopal Hospital. New York
- 1986-1990 Residency in Anatomic and Clinical Pathology, The Mount Sinai Medical Center, New York, NY
- 1990-1991 Fellowship in Surgical Pathology, The Mount Sinai Medical Center, New York, NY
- College of American Pathologists, Fellow and Inspector, Team Leader
- The Joint Commission, Pathologist Specialist and Laboratory Inspector
- American Society for Clinical Pathology, Fellow
- American Association for Clinical Chemistry, Member
- Human Research and Biomedical Investigator – Collaborative Institutional Training Initiative
Certifications and Licenses
- Board Certification by the American Board of Pathology in Anatomic and Clinical Pathology, 1991
- New York State Medical License, 1992
- New York State Laboratory Director Certificate of Qualification including Bacteriology, Blood Banking Collection-Comprehensive, Blood Lead, Blood pH and Gases, Clinical Chemistry, Clinical Toxicology, Cytopathology, Diagnostic Immunology, Endocrinology, Hematology, Histopathology General, Immunohematology, Mycology, Oncology Soluble Tumor Markers, Parasitology, Therapeutic Substance Monitoring/Quantitative Toxicology, Transfusion Services and Virology
Research and Clinical 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 two p53 peptides from its HDM-2-binding domain (i.e., residues 12-26 and 17-26) each attached to a membrane residency peptide (MRP) or pentratin sequence that allows for its transport across the cell membrane. The 12-26-MRP and 17-26-MRP peptides are called PNC-27 and PNC-28, 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 over 40 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. On the other hand, we have tested them against multiple different normal cell lines and have found that they have no effect on cell growth or viability. 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. In high resolution confocal microscopy experiments, we have found that PNC-27 colocalizes with the HDM-2 protein in the cancer cell membrane and that normal cells express little if any HDM-2 in their membranes, explaining the specificity of tumorolytic activity of this peptide. In high resolution immuno scanning electron microscopic experiments on gold-labeled PNC-27 and HDM-2 experiments we have found that PNC-27 and HDM-2 form 1:1 complexes that line transmembrane pores explaining the cytotoxic effect of PNC-27. 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. In further independent experiments it has been found that PNC-27 kills acute myelogenous leukemia stem cells from ten patients transplanted into nude mice with no off-target effects indicating that this peptide can be used effectively to treat and possibly cure this disease. We have shown that PNC-27 also kills non-stem cell AML leukemia cells and solid tissue cancer stem cells including ovarian and colon 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, linked to the MRP sequence, on TUC-3 cells as described above. We found that both peptides induced phenotypic reversion of the TUC-3 cells to normal BMRPA1 cells. On the other hand, treatment of the normal acinar BMRPA1 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.
Supported in part by: NCI RO1, VA Merit Award, Lustgarten Foundation for Pancreatic Cancer Research and the American College of Surgical Oncology.
Space-filling representation of the three-dimensional structure of the anti-cancer peptide, PNC-27, showing the amphipathic nature of the molecule. The green domains show the hydrophobic face of the peptide while the red and blue show the positions of polar negativley and positive charged amino acid residues, respectively on the opposite face of the molecule. The positively and negatively charged residues are seen further to be separated from one another.
- Jayakumar R, Cassai N, Laskar D, Khurram NA, Ladoulis C, Wieczorek R and Pincus MR (2018). Comparison of histopathological diagnosis by whole slide imaging with direct microscopic slide evaluation for a variety of cases. J Appl Clin Path DOI: 10.24983/scitemed.jacp.2018.00065.
- Wang XH, Momeni A, Eid I, Li L, Brackett F, Davis-Smith B Wieczorek R and Pincus MR (2018). Major discrepancies of hcv viral loads determined by real time polymerase chain reaction caused by use of collection tubes that differ only with the presence or absence of serum separator. Ann Clin Lab Sci 48, 286-295.
- Momeni-Boroujeni, A and Pincus MR (2018). Systematic error detection in Laboratory Medicine in Quality Control in Laboratory Medicine, Zaman GS, Ed. TechOpen Publishers, Rijeka, Croatia, Chapter 5, 49-65, I:10.5772/intechopen.72311.
- Chua C, Tifigiu E, Boroujeni A M, Lin B, Laskar D, Shafique K, Zuretti A, Michl J and Pincus MR (2018). Stability of values for the activities of critical enzymes assayed in serum frozen for prolonged time periods. Ann Clin Lab Sci 48, 55-63.
- Carty RP, Lin B, Fridman D and Pincus MR (2018). Low energy conformations for s100 binding peptide from the negative regulatory domain of p53. Prot J, 37, 510-517.
- Lin B, Carty RP and Pincus MR (2020). Low energy conformations for endogenous mu-receptor-specific peptides protein J 39, 217-223. https://doi.org/10.1007/s10930-020-09903.
- Seydafkan S, Michl JP and Pincus MR (2020). Unique features of prostate cancer in african american and west indian patients including diagnosis of high grade cancers using only elevated serum levels of prostate specific antigen (PSA) Ann Clin Lab Sci 50, 55-62.
- Thadi A, Gleeson E, Khalili M, Shaikh MF, Goldstein E, Morano WF, Daniels LM, Grandhi N, Glatthorn H, Richard SD, Cambell PM, Sarafraz-Yazdi E, Pincus MR and Bowne WB. Anti-cancer tumor cell necrosis of epithelial ovarian cancer cell lines depends on high expression of hdm-2 protein in their membranes Ann Clin Lab Sci, in press.
- Thadi A, Lewis L, Goldstein E, Agarwal A, Khaili M, Steele L, Polyak B, Seydafkan S, Bluth MH, Ward KA, Styler M, Campbell PM, Pincus MR and Bowne WB (2020). Targeting membrane hdm-2 by pnc-27 induces necrosis in leukemia cells but not in normal hematopoietic cells Anticancer Res 40, in press. doi:10.21873/anticanres.11xxx.
- Momeni, A., Yousefi, E. and Zuretti, A.: Time series analysis of laboratory values in the context of long-term hospitalized patient mortality. American Journal of Clinical Pathology, doi/10.1093/ajcp/aqy163/5303265, 2019
- Chua, C., Tifigiu, E., Momeni, A., Laskar, D., Shafique, K., Zuretti, A., Michl, J., and Pincus, M.: Stability of Values for the Activities of Critical Enzymes Assayed in Serum Frozen for Prolonged Time Periods. Annals of Clinical & Laboratory Science, 48:618-626 doi: 10.15761/CDP.1000115, 2018
- Laskar, D., Shafique, K., Lu, C. and Zuretti, A.: Evaluation and characterization of monoclonal gammopathies using serum protein electrophoresis in a major urban population: one institution’s experience. Journal of Investigative Medicine, doi: 10.1136/jim-2017-000446, 2017
- Chua, C., Tifigiu, E., Momeni, A., Laskar, D., Shafique, K., Zuretti, A., Michl, J., and Pincus, M.: Stability of values for the activities of critical enzymes assayed in serum. Clinical and Diagnostic Pathology, doi: 10.15761/CDP.1000115, 2017
- Leonardi, G.,Wilson, A. and Zuretti, A.:. Evaluation of respiratory syncytial virus (RSV) direct antigen detection assays for use in point-of-care testing. Journal of Virological Methods, 213, 131-134, 2015
- Leonardi, G.,Wilson, A. and Zuretti, A.: Comparison of Conventional Lateral-flow Assays and a New Fluorescent Immunoassay to Detect Influenza Viruses. Journal of Virological Methods 189, 379-382 Ref: VIRNET12078, 2013
- Gao, L., Lipka, A., Avezbakiyev, B., Zuretti, A and Mustacchia, P.: Synchronous Duodenal Carcinoid and Adenocarcinoma of the Colon. World Journal of Oncology; 3(5):239-242, 2012
- Barnett, W and Zuretti, A.: Recurrent Malignant Chondroid Syringoma. American Journal of Clinical Oncology, Vol 23, No 3, 227-232, 2000
- L. Mesia, and Zuretti, A.: Ossified Intratesticular Mucinous Tumor. Archives of Pathology and Laboratory Medicine; Vol 123, 244-246, 1999
- Cacciarelli, A., Zuretti, A. and Robilotti, J.: Lower Helicobacter Pylori Infection and Peptic Ulcer Disease Prevalence in Patients with AIDS and Suppressed CD4 Counts. American Journal of Gastroenterology; 91, No 9, 1783-1784, 1996
- Cacciarelli,A., Abdel Dayem, H.M and Zuretti, A.: Acute Cholecystitis in AIDS Patients: Correlation of Tc 99M Hepatobiliary Scintigraphy with Histopathological Laboratory Findings and CD counts. The society of Nuclear Medicine, November 1995
- Zuretti, A and Schwartz, I.: Gangrenous Herpetic Whitlow, American Journal of Clinical Pathology; 93:828-830, 1990