Within the first three years of its existence (established May 2006) the Angio Core has accomplished all of the above sited goals and is now expanding their efforts in these individual areas, involving: 1) Collaboration interactions with Dr. David Salomon (NCI/CCR/Mammary Biology and Tumorigenesis Laboratory), who has been working with cripto/apelin/apj involvement in EMT/Breast CA development, has resulted in the identification of a second angiogenic factor encoded in the apelin gene. This new growth factor is generated by the enzymatic processing of apelin-36 giving rise to two distinct peptides;the previously know split-product form the carboxy-terminal region (apelin-13) and a second entity from the amino-terminal region denoted as Selective Apelin-36 Cutting and Amidation peptide (Salcut-NH2). Salcut-NH2 was shown to be a growth stimulator of both blood vessel and lymphatic endothelial cells, mast cells and several anatomically different solid tumor cells. Rabbit antisera to Salcut-NH2 easily identified peptide expression in paraffin section of normal and diseased tissue. A quantitative ELISA was developed that selectively discriminated Salcut-NH2 from apelin-36, having a sensitivity in the sub nanogram/milliliter range. EIR filed with NCI/TTC 2/27/08, TRG Approval 4/9/09, Provisional Patent filed 2/27/09 (61/156, 351), Posted on Federal Register 5/1/09, Possible CRADA interest response from Vivo Biosciences, Inc. 6/11/09, actively involved in Scope-of-Work negotiations. 2) Collaborating with Drs Donald McDonald (USCF) and Brad St.Croix evaluating commercially available neutralizing anti-murine VEGF in several in vivo mouse model system and determining what effect such reagents have on tumor growth and angiogenesis. The Angio Core pre-screened a variety of vendor products said to be effective neutralizer of murine VEGF bioactivity using their established high throughput assay systems and found the R&D goat antisera reagent (Cat#AF-493-NA/Lot No. YU1000812A) to be far superior to other commercial compounds. A large quantity of the R&D reagent was purchase with Angio Core CRADA money and used effectively by Drs. McDonald and St. Croix to block murine-VEGF induced angiogenesis in their animal tumor models. Publications detailing these studies are forthcoming. 3) Development of high throughput assays for the identification of anti-antigenic drugs. Dr. Enrique Zudaire, the groups staff scientist, took the lead on this project and created a fluorescent based, 96-well assay system for detecting drug candidates that blocked endothelial cell proliferation, tube formation or both. Dr. Zudaire used this analytical technology to evaluate the DTP diversity set of small molecule as a Proof-of-Principle approach. Over the past 1.5 yrs and through multiple repeat testing, he has identified several small molecule compounds having anti-angiogenic activity. Recently Dr. Zudaire submitted his finding to the NCI Technology Transfer Center (NCI/TTC) as an EIR, circumventing TRG review via Dr. Robert Wiltrouts signed approval, and resulting in an NCI provisional patent (61/230,667) being filed 7/31/09 thus giving Dr. Zudaire the ability to present his science at the Angiogenesis Gordon Research Conference in Newport, RI (8/2/09 to 8/7/09) without jeopardizing IP status. Dr. Zudaire has chosen seven of his best candidate drugs (that include NSC numbers;19630, 292222, 259969, 122657, 150117, 48300 and 292596 representing 0.4% of the total number of small molecule drugs screened) to do further in vivo characterization studies via DIVAA and nude mouse xenograft testing on low angiogenic and high angiogenic human tumors (50mg drug aliquots received from DTP/Fisher BioServices 8/21/09), and 4) In the initial stages of organizing a angiogenesis/lymphangiogenesis techniques book that expands on the assay systems taught in the Angio Core/FAES sponsored TRAC 28 course given biannually at the NIH/Cloisters. IOS Press has been contacted as a possible publisher and weve met with the chief editor (Yale Altman), who is very supportive of putting this text together. We are now in the process of recruiting recognized experts in the field to write specified chapter regarding in vitro and in vivo model system and negotiating color cost for final publication printings.
