Single-cell Functional Profiling of Lymphocytes for Cancer Immunotherapy

Immunotherapy by harnessing patients’ the immune system has changed the landscape of cancer therapeutics and shown promising and remarkable clinical responses. However, not all the patients would be beneficial from the treatment. Lymphocytes are a significant target in anti-tumor immunotherapy, and the functional assessment of lymphocytes will provide insights on their functional biology and will provide a direct path to the improvement of the treatment efficacy. In the first part of this dissertation, we developed and implemented a methodology based on Timelapse Imaging Microscopy in Nanowell Grids (TIMING) platform that integrates phenotypic profiling and dynamic cytokine secretion with single-cell resolution. Analysis of hundreds of human peripheral nature killer cells (NK cells) suggested that CD56dimCD16+ NK cells are immediate interferon gamma (IFN-γ) secretor upon activation by phorbol 12-myristate 13-acetate (PMA) and ionomycin (< 3 h), and no evidence of cooperation between NK cells to synergistic activation or faster IFN-γ secretion. These results establish our technology as an investigational tool for cellular phenotyping and real-time protein secretion of individual cells in a high-throughput manner and demonstrate that the conventional phenotypic based functional annotation of NK cells might be overly simplistic. In the second part of this dissertation, we performed whole transcriptomic profiling on T cells from acute myeloid leukemia patients (responders and non-responders) who were treated with combination therapy of a hypomethylating agent (5-azacytidine) and an immune checkpoint inhibitor (nivolumab, programmed cell death protein 1/PD-1 inhibitor). Sixty-four patient-derived T cells from peripheral blood or bone marrow (site of disease), which were collected before the initiation of the therapy (baseline, T0) and after the first round of treatment (end of cycle one, EC1), were evaluated. Our results demonstrate (1) treatment-induced gene expression changes on circulating CD8 T cells, and (2) the ratios of effector and exhausted CD8 T cells has the potential to serve as a biomarker for patient stratification.

Name thesis:degree_name

Doctor of Philosophy

Level thesis:degree_level

Doctoral

Discipline thesis:degree_discipline

Chemical Engineering

Grantor

University of Houston

Year dc:date.issued

2019

Author dc:creator

• An, Xingyue

Advisor dc:contributor.advisor

• Varadarajan, Navin

Committee members dc:contributor.committeemember

• Cirino, Patrick C.
• Rimer, Jeffrey D.
• Peng, Weiyi
• Singh, Harjeet

• Cancer Immunotherapy
• Single cell
• Immune checkpoint
• Immunology
• T cells
• PD-1
• Lymphocyte

Statement dc:rights

• The author of this work is the copyright owner. UH Libraries and the Texas Digital Library have their permission to store and provide access to this work. UH Libraries has secured permission to reproduce any and all previously published materials contained in the work. Further transmission, reproduction, or presentation of this work is prohibited except with permission of the author(s).

Language dc:language.iso

eng

Handle dc:identifier.uri

https://hdl.handle.net/10657/5287

OAI identifier oai:identifier

oai:uh-ir.tdl.org:10657/5287