Research
The Gynecologic Oncology Fellowship Program at city Medical Center is one of the oldest fellowship programs in the nation, graduating its first fellow in 1967. The program was site visited in August 2017, and is fully accredited by the ACGME.
The primary goal of this fellowship program is to train competent surgeons and caring physicians who will serve and treat women with gynecological malignancies. These graduates are also expected to be dedicated educators and future clinician-scientists who will further propagate knowledge and interest in this field.
Over the past five decades, this program has made significant contributions to Gynecologic Oncology. This tradition will be continued through the devotion of the faculty and of the graduates.
Josef Michl
Laboratory Space: 2,000 square feet
The laboratory is equipped with tissue culture facilities including two laminar flow hoods, and one laminar flood hood for handling of infectious agents, incubators with and w/o CO2 supply, low, medium and high speed refrigerated centrifuges and several low to medium speed table top centrifuges, rotating and shaking water baths, inverted and standard light and fluorescent photomicroscopes (Zeiss and Nikon, respectively), a Biotek “ELx800” Absorbance Microplate Reader and a Packard “Lumicount” Microplate luminometer both linked to PCs equipped with the reader-specific software and a printer. The laboratory further contains a PCR facility, Omni ultrasonication equipment and chromatography facilities (electrophoresis and immunoblotting) for protein purification including a computer-linked BioRad and Waters HPLC system for peptide and protein isolation as well as instrumentation (gel apparatus, UV illuminator, etc.) for working with DNA & RNA, several refrigerators including a work-in refrigerator for chromatograph, -20oC freezers (one explosion proof, two -80oC freezers and several liquid nitrogen cell storage tanks. Next to each of the 5 workbenches are desk spaces for students and fellows equipped with internet-linked PC computers and telephone. In one of the laboratories a separate small room is the dedicated chemical storage facility for lab-chemicals.
A walk-in cold room (controlled steady 4-6oC) is located on the same floor (4th BSB) as our laboratory. Located in rooms next to the laboratories are institutional core-facilities for immuno-histology, confocal microscopy, which is equipped with a new Olympus confocal laser-scanning microscope FC1000 on an Olympus inverted IX71/IX81 microscope. The microscope is further equipped with a controlled temperature and CO2 long-term incubation chamber enabling observations of live cells. At the same location is the departmental transmission electron microscopy facility (TEM), which is equipped with ultramicrotomes, rapid cell freezing, chemical hood and accessory instruments, and a Zeiss TEM with automatic digital camera linked to PC. On the floor is also located the departmental laboratory for FACS analysis. The facilities are maintained and supervised by dedicated, highly specialized and experienced assistants.
Alan Gintzler
Laboratory Space: 2,500 square feet
Biochemistry: Chromatography refrigerator; 3-80oC freezers; 3 power supplies for gel chromatography 2 Sorvall RC-5B (super speed); Beckman L2-65B (ultracentrifuge); 1 Beckman table top untra-centrifuge; 2 platform shakers (Eberbach); HPLC system (Beckman) consisting of two pumps (ALTEC 110A) gradient liquid chromatograph model 332, microprocessor (ALTEX, model 420), Altex 210 sample injector, Rainin 7010 sample injector,) spectra/glo filter fluorometer, HPLC accessories such as) C18 reverse phase columns, assortment of different size injection loops, Eldex pumps to be used with an on line precolumn concentrator of for postcolumn derivatization for fluorescent detection of peptides); Beckman System Gold HPLC system with System Gold 126 Solvent Module and 168 detector with SS 420X software fraction collectors, (LKB model 2112-redirac); Brandel Cell harvestor; multiple analytical and top-loading balances: Waters absorbance Detector, Model 441 Harvard trip balance (Ohaus); (1) Ultrasonic cleaner (Branson, model 92); shaking Incubators: (1) Labline, (1) Eberbach; (1) tissue chopper; Homogenizers: (1) polytron (Brinkman) digital pH meter: 2 precision direct drive high vacuum pump, 3 fume hoods; assorted accessories for biochemistry such as hot plate stirrers, repeat pipets, vortex mixer etc. Tissue culture facilities include two rooms equipped with a liquid nitrogen freezer, laminar flow exhaust hoods (Forma Scientific) and CO2 incubators (maintained at 10% and 8% CO2; Forma Scientific); PerkinElmer micro-beta radioactive plate reader; fluorescent plate reader (Envision, PerkinElmer;); CCD camera (Gene Gnome, Syngene)
Physiology: (2) Stimulators (Grass, S48); (1) crown DC300A amplifier; (1) bipolar generator (custom built); (2) temperature water circulators; (2) peristaltic pumps (LKB, 2132 microperpex); (2) polygraphs (Gould, model 2200 and 220-; (1) isometric force transducer (Grass, FT-03); (1) isotonic transducer (Harvard); assortment of organ bath and tissue superfusion chambers, (2) refrigerators with -20oC freezers; (1) Revco, -70oC freezer (ULTRA LOW); Microscopy: (1) dissecting scope (WILD; (1) fiberoptics light source (VOLPI)
Josef Michl & Ehsan Yazdi
Anti-Cancer Therapeutic and Drug Design
Ideally, anticancer medicines would kill cancer cells but leave normal cells alone. Thus far, anti-cancer agents cause the death of both cancer and normal cells, causing major deleterious side effects. Now, our team of researchers at city Medical Center has discovered several small peptides that meet this ideal, i.e., cause cancer cell death but do not affect normal cells. These peptides, called PNC-27 and PNC-28, were originally designed to block the interaction of p53 with a protein that causes its breakdown. In so doing, cancer cells would die by apoptosis or programmed cell death.
Our inventions encompass sets of peptides from the p53 anti-onogene protein. We have designed these peptides from p53 protein using computer-based molecular modeling of the three-dimensional structures of this protein. 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. Currently, we are investigating the use of this anti-cancer peptide in treating ovarian cancer in a syngeneic mouse model.
Two recent publications demonstrate the PNC-27 Mechanism of Action:
-
Sarafraz-Yazdi E, Bowne WB, Adler V, Sookraj KA, Wu V, Shteyler V, Patel H, Oxbury W, Brandt-Rauf P, Zenilman ME, Michl J, Pincus MR. Anticancer peptide PNC-27 adopts an HDM-2-binding conformation and kills cancer cells by binding to HDM-2 in their membranes. Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):1918-23. Epub 2010 Jan 11.
-
Sookraj KA, Bowne WB, Adler V, Sarafraz-Yazdi E, Michl J, Pincus MR. The anti-cancer peptide, PNC-27, induces tumor cell lysis as the intact peptide. Cancer Chemother Pharmacol. 2010 Feb 25. [Epub ahead of print]