Author ORCID Identifier
Date of Graduation
12-2021
Document Type
Dissertation (PhD)
Program Affiliation
Biomedical Sciences
Degree Name
Doctor of Philosophy (PhD)
Advisor/Committee Chair
Prahlad Ram, Ph.D.
Committee Member
Faye Johnson, M.D., Ph.D.
Committee Member
Ju-Seog Lee, Ph.D.
Committee Member
Rick Wetsel, Ph.D.
Committee Member
Nidhi Sahni, Ph.D.
Committee Member
Bill Mattox, Ph.D.
Abstract
Metastatic lung cancer has a 5-year survival rate of 5%. Lung cancers tend to be asymptomatic until late stages, and almost 90% are not diagnosed until they are advanced. Metastases are very rare events, often initiated by a single cell from a primary tumor into a new niche at a distant location. Investigation of the early metastatic process is of urgent need for the development of early diagnostics and targeted therapeutics. We performed a proof-of-concept CRISPR/Cas9 whole genome knockout screen in the A549 lung adenocarcinoma cell line and utilized a novel ex vivo 4D lung metastasis model to find gene candidates to test in vitro. I prepared samples and performed next generation sequencing. I created a computational pipeline to demultiplex samples and align reads to the library. I recovered genes previously implicated in lung cancer and metastasis enriched in the Primary and Metastasis samples. SPI1 was identified as the central node in network analysis of the Primary and Metastases and present in every sample. SPI1 was validated in vitro as a novel transcription factor involved in lung cancer metastasis.
Keywords
Metastasis, Lung Cancer, SPI1, CRISPR Screen, 4D Lung Model
Included in
Cancer Biology Commons, Cell Biology Commons, Disease Modeling Commons, Laboratory and Basic Science Research Commons