Hao Chen

Analytical Development 1. Development of analytical methods to characterize the purity, stability, phenotype and function of CAR-T drug products and viral vectors. 2. Act as Subject Matter Expert for multiple assays: IncuCyte based cytotoxicity and proliferation; multichannel Flow cytometry-based purity, cell health, phenotype and activation; multiplex based bulk cytokine assays (MSD, Ella, Luminex…); qPCR and ddPCR based assays, including VCN, AAV genome titer and sLVV DLTA. 3. Method qualification and phase appropriate validation. 4. Critical reagents generation and qualification: assay controls (cells and virus), antibodies, ddPCR positive controls… 5. SOP writing and transferring to downstream teams. 6. IND enabling and submission. Responded to reviewer’s comments timely and persuasively to accelerate approval. 7. Exploring novel hypothesis/ideas to develop in vitro models and methods with functional relevance and predictive indications: a. Chronic/repeated stimulation model to characterize persistent potency. b. Single cell sequencing and data analysis: Discover specific functional relevant biomarkers. c. Multiplex model: simultaneously analyzing cytotoxicity, proliferation, differentiation within one assay.

Plasmid production. Supervision on all the stages of molecular cloning. Although only worked in this role for one month, I was able to find critical issues on SOPs and operations, designed and executed multiple studies/experiments to figure out the root causes, leading to significant improvement of the successful rate.

TCR-T early development and candidate selection. Generation of specific anti-tumor T cells from PBMC with lentivirus transduction. In vitro evaluation of the engineered T cell candidates: tetramer binding; peptide titration; tumor cell co-culture; Flow cytometry for T cell activation markers; ELISA for cytokine induction; cross-activity screening; xCELLigence cytolytic assays. In vivo evaluation of the candidates: generated tumor xenograft models with NSG and NCG mice. Injection of engineered T cells into mice and monitoring tumor growth; multichannel flow cytometry analysis on the circulating and tumor infiltrating T cells; Data analysis and visualization for presentation at R&D and PD team meetings.

Single cell RNA sequencing with human Desmoplastic Small Round Cell Tumor (DSRCT) and the adjacent omental tissues. Dissected the intratumor heterogeneity and discovered the novel cancer stem cells as well as the unique tumor microenvironment, including NKT cells and mesenchymal cells only presented in tumor tissues. Identified specific cell surface markers on the cancer stem cells and the tumor associated immune and stromal cell populations.

Utilizing homogeneous recombination, tagged collagen 1A2 gene with EGFP reporter in zebrafish to study the dynamic behaviors of fibroblasts in skin regeneration and repair. Mastering high resolution confocal microscopy in tracing the spatiotemporal live cell movements during wound healing and skin regeneration.

Interrogation on the molecular mechanism of esophageal development and diseases, using mouse and zebrafish models. Identified a couple of biomarkers and potential therapeutic target genes/pathways in esophageal cancers, including Sox2, Pax9, Cdx2, NFkB and Nrf2/Keap1.

Discovery and cloning of a novel liver specific estrogen responsive gene (Vtg1) and the corresponding enhancer sequence. Generated an estrogen responsive transgenic zebrafish line in which the EGFP expression was driven by the promoter of the Vtg1 gene. This animal model was used for studies on liver development and diseases, as well as detecting environment estrogens.

Molecular construction of a soluble peptide-HLA complex to activate tumor antigen specific cytotoxic T cells in vitro and in vivo.