Author ORCID Identifier


Date of Graduation


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.


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.


Metastasis, Lung Cancer, SPI1, CRISPR Screen, 4D Lung Model