• Overall goal: Improve manufacturing and clinical efficacy for adoptive T-cell therapies against solid tumors by genetically modifying T-cells to express the synthetic coreceptor CD8α:MyD88.• Sub-aims: (1) Investigate how MyD88 signaling in CD8 T-cells increases sensitivity to IL-2 to achieve improved manufacturing capabilities and reduced IL-2 mediated toxicities associated with adoptive T-cell therapies. (2) Determine the role of MyD88 signaling in promoting CD8+ T-cell endoplasmic reticular homeostasis to enhance antitumor immunity.• Efforts around my thesis project contributed to the incorporation of startup: TrAMPoline Pharma and a $10M award to support phase 1 clinical trials for CD8α:MyD88 TIL therapy.• Advisor: Dr. Eduardo DavilaTechniques employed: • Viral Transduction• Transfection• 8+ Flow Cytometry• FACS• ELISA/Western Blot/Protein Arrays• Multiplex IF/IHC• Light/Confocal microscopy• In-vivo syngeneic mouse tumor models for adoptive T-cell therapies (Pmel/B16, OT1/OVA)• Omics: Bulk and single-cell RNAseq, Proteomics, Metabolomics, ATACseq• R Programming• Recombinant DNA Technology• Nucleic Acid/Protein Isolation and Quantification• ChIP-PCR, RT-qPCR, primer design• Primary mouse/human immune cell isolation and culture• Cytotoxicity assays• Cytokine/chemokine quantification• Phenotypic characterization• Proliferation assessment• Effector-target co-culture assays• Metabolic fitness assays (seahorse, Mitotracker, TMRM/TMRE)• Open bioreactor cell culture systems (GREX)• Potency Assays• Stable cell line generation

My role as senior venture fellow is to assist CU Innovations and Gates Institute at the University of Colorado Anschutz Medical Campus (CU-AMC) in streamlining the commercialization of regenerative technologies, and cell and gene therapies that originate from research labs on campus. To do so, I perform the following duties:1) Assist the CU Innovations licensing and business development teams with innovation review and commercialization activities. Examples include:- Support due diligence analyses describing value proposition and distinct technological and market advantages of cell and gene therapy/regenerative medicine technologies shared from the CU-AMC community.- Meet and interview investigators with licensing managers to develop IP protection strategies- Develop marketing strategies for innovation portfolio including: finding potential commercial partners, identifying key partners and resources for development, making initial contact with potential partners, and sharing marketing campaigns + tracking progress2) Support management of funding programs designed to lower the barrier to commercialization for inventors from academic institutions: Startup Toolbox and Gates Institute Grubstake Awards. Examples include:- Developing marketing materials and dissemination of information to attract applicants + managing web portal content- Serving as initial point of contact for the academic community for queries relating to application process- Evaluate resource needs of applicants and connect them to the appropriate resources before and after the application process- Coordinate with Grubstake Fund management team and Scientific Investment Advisory Committee to review applications (includes being the point person to describe the scientific background for each application)

Supporting systems engineering team with R&D and V&V testing for rapid, miniaturized, and portable PCR diagnostics

Lead in development, optimization, and training of various reagent manufacturing processes and their respective quality control methods in a cGMP setting.Collaborated with QA/QC team to successfully procure ISO 13483:2016 certification and to perform process and method validations leading to successful entry into clinical trials and analytical studies for FDA 510(k) submission for the Click Sexual Health Test. Lead in the R&D feasibility studies to develop a rapid nucleic acid based assay to detect antimicrobial resistance (AMR) in Neisseria gonorrhoeae (NG) that is compatible with the Click Sexual Health Test platform. Research pertaining to AMR NG project was submitted and awarded for the following grants: NIH funded AMR Rapid, Point-of-Need Diagnostic Test Challenge step 2 and the Small Business Innovation Research (SBIR) grant.Lead in R&D studies to develop an instrument-free rapid blood sample preparation method to extract viral RNA with Click Sexual Health Test platform compatibility in collaboration with USAMRIID and the NRL. I led weekly meetings with our collaborators to discuss results and form strategic next steps to ensure mission alignment. Lead in the design and testing of surface-bound capture oligos for the detection of influenza A, influenza B, and the bacteriophage MS2 for the development of the Visby Respiratory Panel. Lead in design of lab floorplans and equipment layout for lab facility relocation

- Key member in supporting cross functional teams in clinical, quality control, and manufacturing.- Established and maintained inventory for clinical specimens. - Lead in multiple PCR analytical development studies including, but not limited to, incoming inspection and pre/post-lyophilization assays for Taq DNA Polymerase enzyme activity, and incoming inspection assays for PCR primers and dNTPs- Lead in preclinical vaginal and target infectious organism(s) sample stability studies- Lead in preclinical manufacturing process reagent stability studies- Lead in preliminary preclinical benchtop R&D LOD, cross reactivity, and interfering substances testing- Developed initial framework for a molecular based microbial enumeration method to achieve more precise and accurate quantitation of bacterial cells as compared to the current gold standard: visual counting.- Developed, authored and optimized manufacturing processes and QC methods- Authored various other quality documents such as PS's, EQ's, process deviations, test protocols, test reports, supplier evaluations, NMRs, etc- Collaborated with manufacturing team for reagent process transfer/scale up- Presented experimental data to cross functional teams (R&D, manufacturing, quality, upper management) to drive key decisions- Basic understanding of regulations such as FDA, cGMP, GLP, OSHA, GDP and ISO certifications- Collaborated with QA to prepare and maintain labs/prepare documentation according to GLP/GMP for Phase I ISO 13485 audit- Key member in the successful completion of various QC Method Validations and Process Validations (including execution, development, and documentation of process deviations/LHRs)- Proficient in the following laboratory techniques: Lyophilization, pipetting, qPCR, ddPCR, DNA isolation, microbial cell culture, gel electrophoresis (agarose and bioanalyzer using DNA kit)

Duties included:-Clinical, microbial, chemical, equipment and lab consumable inventory management-Lead in preclinical analytical and stability studies-Key member in orchestrating relocation and organization of new lab facilities (Includes R&D and GMP labs). Moving to a site that's 10 times the size of the former facility -Independently planned and executed vital experiments for the development of proprietary assay components, including, but not limited to, lyophilized PCR reagent mixes, collection media, and DNA extraction protocol/reagents-Independently analyzed experimental results and presented them at team meetings with upper management- Collaborated with cross-functional teams of engineers and QA/RA experts to develop key components in the product-Performed the following experimental techniques on a daily basis: DNA extraction (dirty and clean prep), gel electrophoresis, qPCR -Followed and supported development of manufacturing SOP's, QC procedures and other QMS documentation

Designed and conducted experiments to investigate genomic rearrangements in Tetrahymena Thermophila using:-DNA recombinant technology-PCR-DNA electrophoresis-ciliate genome database 15 hrs/wk

Coordinated experiments with the purpose of developing a novel method to analyze gene expression profiles from 3-D cell cultures at the single-cell level using:-light microscopy-formalin fixation-DNA/RNA isolation and quantification-immunoblot assays, and RT-qPCRSpecifically worked with 3-D cultures modeling Barrett's Esophagus and that of esophageal adenocarcinomas.

Optimized conditions for RNA extraction without the use of a wash column to reduce lab costs.Performed experiments with emphasis on gene expression profiles pertaining to the progression of esophageal adenocarcinoma from Barrett's Esophagus at the single-cell level using RT-qPCR and immunoblot assays.