SBIR-STTR Award

Despite progress in immunotherapies, there are significant challenges to overcome to make them more broadly applicable to different cancers and more effective for patients. However, current efforts to improve adoptive cell therapies and immune checkpoint blockade as well as cancer vaccines are hampered by the lack of a method to characterize the functional and phenotypical changes of different subpopulations of T cells over time at high throughput. The goal of this SBIR Phase II project is to develop a novel flow-cytometry method that can solve this technological bottleneck. The time-lapse flow cytometry uses laser particle (LP) barcodes to track each cell across flow measurements taken at different time points. Built on demonstrated proof-of-concept data, this project is focused on establishing a set of novel assays for characterizing T cells in response to different stimuli in a time-resolved manner. The first specific aim is to demonstrate short-term time-lapse assays to profile cytokine secretion (t-SEC). Secretion of TNF, IFNγ and IL10 from millions of the same cells in response to different types of stimulation are measured at multiple timepoints over 24 hours. The second specific aim is to demonstrate long-term time-lapse assay to monitor phenotype (t-PHENO), including T-cell exhaustion over 10 days. The third specific aim is to validate the t-SEC and t-PHENO assays with patient samples. The time- resolved, high-throughput, high-parameter assays are expected to accelerate the development of more effective and durable immunotherapies for cancer (>$100 B global market). Beyond immunotherapy, time-lapse flow cytometry is also expected to be useful in fundamental immunological research and vaccine development.

Public Health Relevance Statement:
Narrative This project aims to foster fundamental discoveries and innovative research for the improvement of immunotherapies to treat cancer by harnessing the patients' own immune cells. A major technological limitation is the lack of methods to characterize immune cells over time at high- throughput. The proposed research to develop novel time-lapse flow cytometry addresses this limitation, accelerating immunology and immune-oncology research and development.

Project Terms:
Acceleration; Antibodies; Bar Codes; barcode; Biological Assay; Assay; Bioassay; Biologic Assays; Malignant Neoplasms; Cancers; Malignant Tumor; malignancy; neoplasm/cancer; Cell physiology; Cell Function; Cell Process; Cellular Function; Cellular Physiology; Cellular Process; Subcellular Process; Cell Separation; Cell Isolation; Cell Segregation; Cell Separation Technology; cell sorting; Cells; Cell Body; Enzyme-Linked Immunosorbent Assay; ELISA; enzyme linked immunoassay; Exhibits; Flow Cytometry; Flow Cytofluorometries; Flow Cytofluorometry; Flow Microfluorimetry; Flow Microfluorometry; flow cytophotometry; Goals; Human; Modern Man; Immunotherapy; Immune mediated therapy; Immunologically Directed Therapy; immune therapeutic approach; immune therapeutic interventions; immune therapeutic regimens; immune therapeutic strategy; immune therapy; immune-based therapies; immune-based treatments; immuno therapy; instrumentation; Interferon Type II; IFN-Gamma; IFN-g; IFN-γ; IFNG; IFNγ; Immune Interferon; Interferon Gamma; lFN-Gamma; Interleukin-2; Co-Stimulator; Costimulator; Epidermal Thymocyte Activating Factor; IL-2; IL2 Protein; Interleukin 2; Interleukin 2 Precursor; Interleukin II; Interleukine 2; Interleukine 2 Precursor; Interleukine II; Lymphocyte Mitogenic Factor; Mitogenic Factor; T cell growth factor; T-Cell Growth Factor; T-Cell Stimulating Factor; Thymocyte Stimulating Factor; Kinetics; Lasers; Laser Electromagnetic; Laser Radiation; Marketing; Methods; Patients; Phenotype; Production; Quality Control; Research; research and development; Development and Research; R & D; R&D; Research Personnel; Investigators; Researchers; Specificity; Standardization; T-Lymphocyte; T-Cells; thymus derived lymphocyte; Technology; Time; Treatment Protocols; Treatment Regimen; Treatment Schedule; cytokine; Measures; CSIF; CSIF-10; Cytokine Synthesis Inhibitory Factor; IL-10; IL10; IL10A; Interleukin 10 Precursor; Interleukin-10; Treatment outcome; improved; Chronic; Phase; Chemicals; Stimulus; Individual; Fostering; Measurement; anti-cancer immunotherapy; anticancer immunotherapy; immune-based cancer therapies; immunotherapy for cancer; immunotherapy of cancer; cancer immunotherapy; Collaborations; cancer cell; Malignant Cell; Adoptive Cell Transfers; adoptive cell therapy; adoptive cellular therapy; Cancer Vaccines; Antineoplastic Vaccine; Neoplasm Vaccines; Tumor Vaccines; anti-tumor vaccine; antitumor vaccine; vaccine for cancer; exhaustion; Scientist; Hour; Immune; Immunes; Protocols documentation; Protocol; particle; Performance; vaccine development; develop a vaccine; develop vaccines; development of a vaccine; Peripheral Blood Mononuclear Cell; PBMC; novel; Cell surface; Reporting; TNF gene; (TNF)-a; Cachectin; Macrophage-Derived TNF; Monocyte-Derived TNF; TNF; TNF A; TNF Alpha; TNF-a; TNFA; TNFa; Tumor Necrosis Factor; Tumor Necrosis Factor-alpha; Sampling; response; programmed cell death protein 1; PD 1; PD-1; PD1; programmed cell death 1; programmed death 1; sle2; systemic lupus erythematosus susceptibility 2; Address; Aliquot; Contractor; Data; Detection; Immunooncology; immune-oncology; immuno oncology; immunology oncology; oncoimmunology; Prediction of Response to Therapy; predict therapeutic response; predict therapy response; predict treatment response; therapy prediction; treatment prediction; treatment response prediction; Resolution; resolutions; in vitro Assay; Cancer Patient; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Validation; validations; Immunologics; Immunochemical Immunologic; Immunologic; Immunological; Immunologically; Monitor; Process; Development; developmental; cellular imaging; cell imaging; innovate; innovative; innovation; resistant; Resistance; commercialization; bio-markers; biologic marker; biomarker; Biological Markers; T cell response; customized therapy; customized treatment; individualized patient treatment; individualized therapeutic strategy; individualized therapy; individualized treatment; patient specific therapies; patient specific treatment; tailored medical treatment; tailored therapy; tailored treatment; unique treatment; individualized medicine; lab development; laboratory development; phenotypic marker; phenotypic biomarker; systemic toxicity; check point blockade; checkpoint blockade; immune check point blockade; immune checkpoint blockade