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.
","abstract_html":"<p>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.<strong> </strong>We performed a proof-of-concept CRISPR/Cas9 whole genome knockout screen in the A549 lung adenocarcinoma cell line and utilized a novel <em>ex vivo 4D </em>lung metastasis model to find gene candidates to test <em>in vitro. </em>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 <em>in vitro</em> as a novel transcription factor involved in lung cancer metastasis.</p>","abstract_has_math":false,"creators":["Plumer, Alexandria",""],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation (PhD)","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Prahlad Ram, Ph.D.","Faye Johnson, M.D., Ph.D.","Ju-Seog Lee, Ph.D."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2021,"date_issued":"2021-12-01T08:00:00Z","date_published":"2021-12-01T08:00:00Z","updated_at":"2026-07-24T05:49:54Z","subjects":["Metastasis","Lung Cancer","SPI1","CRISPR Screen","4D Lung Model","Cancer Biology","Cell Biology","Disease Modeling","Laboratory and Basic Science Research","Medicine and Health Sciences"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1146","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Prahlad Ram, Ph.D.","Faye Johnson, M.D., Ph.D.","Ju-Seog Lee, Ph.D."]},{"key":"dc:creator","label":"Author","values":["Plumer, Alexandria",""]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2021-10-08T07:00:00Z"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation (PhD)"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Metastasis","Lung Cancer","SPI1","CRISPR Screen","4D Lung Model","Cancer Biology","Cell Biology","Disease Modeling","Laboratory and Basic Science Research","Medicine and Health Sciences"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1146"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["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.
"]},{"key":"dc:title","label":"Title","values":["4D Ex Vivo Crispr/Cas9 Whole-Genome Screen to Identify Genes Regulating Lung Cancer Metastasis"]}]}],"canonical_facts":{"dc:contributor":["Prahlad Ram, Ph.D.","Faye Johnson, M.D., Ph.D.","Ju-Seog Lee, Ph.D."],"dc:creator":["Plumer, Alexandria",""],"dc:date.available":["2021-10-08T07:00:00Z"],"dc:description.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.
"],"dc:identifier":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/1146"],"dc:subject":["Metastasis","Lung Cancer","SPI1","CRISPR Screen","4D Lung Model","Cancer Biology","Cell Biology","Disease Modeling","Laboratory and Basic Science Research","Medicine and Health Sciences"],"dc:title":["4D Ex Vivo Crispr/Cas9 Whole-Genome Screen to Identify Genes Regulating Lung Cancer Metastasis"],"thesis:degree_level":["Dissertation (PhD)"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T05:49:54Z"}