Coordinated with industry partners for project updates and direction. Generated and optimized in vitro screening assays for activity of a previously unpublished immune-oncology myeloid protein. Leveraged scRNAseq, WB, qRT-PCR and LC/MS/MS to elucidate correct readout targets and conditions. Characterized functional effect of novel protein deletion on FACS isolated intratumoral monocytes and macrophages.

Led three independent myeloid cell immune-oncology research projects. 1) Investigating the transcriptional profiles of intratumoral monocytes and macrophages rapidly isolated from primary untreated human clear cell renal cell carcinoma (ccRCC) specimens, 2) elucidating the role of PI3Kgamma in immunosuppressive tumor associated macrophages in prostate cancer, 3) investigation of the mechanistic role of a previously unpublished molecule expressed on myeloid cells and its immunotherapeutic potential across multiple preclinical models of cancer.Utilized small molecule inhibitors against PI3Kgamma in vitro to screen for phenotypic differences. Performed flow cytometry, western blotting, and qRT-PCR to interrogate phenotypic alterations. Utilized multiple in vivo genetic knockout models and pharmacological inhibition models (oral gavage, intraperitoneal injections, subcutaneous injection) to test the effects of disrupting various protein signaling pathways on tumor growth. Performed extensive single cell RNA sequencing experiments to investigate the transcriptional alterations after genetic deletion of the novel molecule. Capitalized on the existing RNAseq analysis platform Ingenuity Pathway Analysis (IPA) to produce hypothesis generating data. These data identify testable putative transcription factors and pathways controlled by this novel molecule and are being leveraged to generate in vitro screening assays for therapeutic molecules. Utilized LC/MS/MS to successfully test specific hypotheses generated from the scRNAseq data analysis. Coordinated with industry partners to generate and test specific antibody clones targeting the novel molecule. Using molecular cloning techniques, generated an in vitro proximity ligation assay (PLA) cell line model to screen for cytoplasmic interacting partners after novel protein ligation. Trained and managed 5 rotation students, conceptualizing, and leading the experimental designs of multiple scientific projects.

Coordinated and led a weekly small group session, focused on discussing primary research articles which underlay a wide array of immunology concepts and techniques described in the lecture portion of the class. Led review sessions, held office hours, created exam questions and graded exams.

Coordinated and led a weekly small group session, focused on discussing primary research articles which underlay a wide array of immunology concepts and techniques described in the lecture portion of the class. Led review sessions, held office hours, created exam questions and graded exams.

Developed and optimized a standard operating procedure (SOP) for rapid FACS isolation of lymphocytes from fresh primary human peripheral blood and tumor specimens. This SOP allowed isolation of pure intratumoral lymphocytes within 4-5 hours of clinical resection of tumor tissue, through strong coordination and communication across multiple healthcare and research teams. Generated and optimized a 9-color flow cytometry panel to isolate primary lymphocytes from clinical patient specimens. This panel focused on identifying naïve and effector CD8+ T cells, and naïve, effector and regulatory CD4+ T cells. Optimized direct FACS sorting into lysis buffer for RNA extraction for very low (>20,000) cell counts. Was able to successfully and reproducibly isolate RNAseq quality RNA from samples as low as ~500 cells. Developed and optimized three 8-10 color flow panels for phenotyping lymphocytes from peripheral and tumor sites from samples obtained from clinical trial specimens. These panels focused on 1) Surface markers, 2) Transcription Factors, 3) Cytokine production.Isolated FFPE RNA from archived human tissue sources for Nanostring analysis. Performed in vitro cell culture experiments testing the effect of inflammatory cytokines on prostate cancer cell phenotypes. Used the Small Animal Radiation Research Platform (SARRP) to study the effect of focal brain irradiation on circulating immune cells. Determined the absolute cell counts of peripheral immune cells using flow cytometry cell counting bead kit. Performed ELISA’s to determine cytokine alterations in lymph nodes after radiation. Maintained an animal colony of >30 strains. Weaned, genotyped (PCR, flow cytometry) and bred each strain successfully for 4 years before transitioning into a Ph.D. training program. Organized and maintained laboratory reagent and consumables stocks, animal protocols, new student on-boarding procedures, and monitored funding utilization.