{"id":{"repo_id":"utswmed","oai_identifier":"oai:utswmed-ir.tdl.org:2152.5/10804"},"canonical_url":"https://search.dev.ndltd.org/etd/utswmed/oai:utswmed-ir.tdl.org:2152.5/10804","repository":{"repo_id":"utswmed","name":"University of Texas Southwestern Medical Center","base_url":"https://utswmed-ir.tdl.org/server/oai/request"},"display":{"title":"Consequences of Pathogenic Mitochondrial DNA Mutations in Cancer Progression","abstract":"Mitochondrial DNA (mtDNA) mutations are prevalent in human cancers; however, their functional impact on cancer progression remains controversial and uncertain. This uncertainty is primarily due to a lack of suitable models for controlled functional studies. This thesis addresses this limitation by developing a novel protocol for the creation of nuclear isogenic transmitochondrial cancer cell lines with defined pathogenic mutations, and secondly, by the establishment of a collection of patient tumor xenograft models with varied mtDNA status. Chapter two details the development of a new cytoplasmic hybrid (cybrid) generation method that simplifies the creation of transmitochondrial cybrids and enables transplantation of any mtDNA within any cell line. Utilizing this protocol, we developed isogenic melanoma lines with targeted mtDNA mutations. In chapter three, these melanoma cybrids are characterized in vivo, revealing that oxidative phosphorylation dysfunction does not block tumor growth. However, mtDNA mutations were found to significantly hinder spontaneous metastasis, largely due to impaired migration and invasion at metabolically restrictive conditions. Additionally, analysis of selective pressures exerted on the mitochondrial genomes in heteroplasmic cybrid cells demonstrated that tumor growth suppressed the pathogenic variant allele frequency (VAF) of mtDNA mutations. Lastly, chapter four analyzes renal cell carcinoma (RCC) patient-derived xenografts from the UT Southwestern Kidney Program, identifying RCC lines with either wildtype or pathogenic mtDNA mutations in Complex I subunits. RCC mtDNA mutations were validated through genetic and functional analyses. Additionally, it was found that RCC cancers with mtDNA mutations were associated with altered transcriptional statuses. Collectively, this thesis elucidates the relationship between mtDNA mutations and tumor development, employs innovative methods to examine mitochondrial function, and underscores the importance of mitochondrial genetics in oncology.","abstract_html":"Mitochondrial DNA (mtDNA) mutations are prevalent in human cancers; however, their functional impact on cancer progression remains controversial and uncertain. This uncertainty is primarily due to a lack of suitable models for controlled functional studies. This thesis addresses this limitation by developing a novel protocol for the creation of nuclear isogenic transmitochondrial cancer cell lines with defined pathogenic mutations, and secondly, by the establishment of a collection of patient tumor xenograft models with varied mtDNA status. Chapter two details the development of a new cytoplasmic hybrid (cybrid) generation method that simplifies the creation of transmitochondrial cybrids and enables transplantation of any mtDNA within any cell line. Utilizing this protocol, we developed isogenic melanoma lines with targeted mtDNA mutations. In chapter three, these melanoma cybrids are characterized in vivo, revealing that oxidative phosphorylation dysfunction does not block tumor growth. However, mtDNA mutations were found to significantly hinder spontaneous metastasis, largely due to impaired migration and invasion at metabolically restrictive conditions. Additionally, analysis of selective pressures exerted on the mitochondrial genomes in heteroplasmic cybrid cells demonstrated that tumor growth suppressed the pathogenic variant allele frequency (VAF) of mtDNA mutations. Lastly, chapter four analyzes renal cell carcinoma (RCC) patient-derived xenografts from the UT Southwestern Kidney Program, identifying RCC lines with either wildtype or pathogenic mtDNA mutations in Complex I subunits. RCC mtDNA mutations were validated through genetic and functional analyses. Additionally, it was found that RCC cancers with mtDNA mutations were associated with altered transcriptional statuses. Collectively, this thesis elucidates the relationship between mtDNA mutations and tumor development, employs innovative methods to examine mitochondrial function, and underscores the importance of mitochondrial genetics in oncology.","abstract_has_math":false,"creators":["Shelton, Spencer Dewayne"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Whitehurst, Angelique Wright","Morrison, Sean J.","McFadden, David G.","Mishra, Prashant"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2026,"date_issued":"2026-06-15T19:42:26Z","date_published":"2026-06-15T19:42:26Z","updated_at":"2026-07-24T05:52:08Z","subjects":["DNA, Mitochondrial","Melanoma","Mutation","Neoplasm Metastasis"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["1596185292"],"render_values":[{"text":"1596185292","href":null,"code":true}]}]},"links":{"outbound_url":"https://hdl.handle.net/2152.5/10804","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Whitehurst, Angelique Wright","Morrison, Sean J.","McFadden, David G.","Mishra, Prashant"]},{"key":"dc:creator","label":"Author","values":["Shelton, Spencer Dewayne"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2026-06-15T19:42:26Z","2024-05","May 2024","2026-06-15T19:42:27Z"]},{"key":"dc:type","label":"Dc Type","values":["Thesis","text"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["DNA, Mitochondrial","Melanoma","Mutation","Neoplasm Metastasis"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://hdl.handle.net/2152.5/10804","1596185292"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Mitochondrial DNA (mtDNA) mutations are prevalent in human cancers; however, their functional impact on cancer progression remains controversial and uncertain. This uncertainty is primarily due to a lack of suitable models for controlled functional studies. This thesis addresses this limitation by developing a novel protocol for the creation of nuclear isogenic transmitochondrial cancer cell lines with defined pathogenic mutations, and secondly, by the establishment of a collection of patient tumor xenograft models with varied mtDNA status. Chapter two details the development of a new cytoplasmic hybrid (cybrid) generation method that simplifies the creation of transmitochondrial cybrids and enables transplantation of any mtDNA within any cell line. Utilizing this protocol, we developed isogenic melanoma lines with targeted mtDNA mutations. In chapter three, these melanoma cybrids are characterized in vivo, revealing that oxidative phosphorylation dysfunction does not block tumor growth. However, mtDNA mutations were found to significantly hinder spontaneous metastasis, largely due to impaired migration and invasion at metabolically restrictive conditions. Additionally, analysis of selective pressures exerted on the mitochondrial genomes in heteroplasmic cybrid cells demonstrated that tumor growth suppressed the pathogenic variant allele frequency (VAF) of mtDNA mutations. Lastly, chapter four analyzes renal cell carcinoma (RCC) patient-derived xenografts from the UT Southwestern Kidney Program, identifying RCC lines with either wildtype or pathogenic mtDNA mutations in Complex I subunits. RCC mtDNA mutations were validated through genetic and functional analyses. Additionally, it was found that RCC cancers with mtDNA mutations were associated with altered transcriptional statuses. Collectively, this thesis elucidates the relationship between mtDNA mutations and tumor development, employs innovative methods to examine mitochondrial function, and underscores the importance of mitochondrial genetics in oncology."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Consequences of Pathogenic Mitochondrial DNA Mutations in Cancer Progression"]}]}],"canonical_facts":{"dc:contributor":["Whitehurst, Angelique Wright","Morrison, Sean J.","McFadden, David G.","Mishra, Prashant"],"dc:creator":["Shelton, Spencer Dewayne"],"dc:date":["2026-06-15T19:42:26Z","2024-05","May 2024","2026-06-15T19:42:27Z"],"dc:description":["Mitochondrial DNA (mtDNA) mutations are prevalent in human cancers; however, their functional impact on cancer progression remains controversial and uncertain. This uncertainty is primarily due to a lack of suitable models for controlled functional studies. This thesis addresses this limitation by developing a novel protocol for the creation of nuclear isogenic transmitochondrial cancer cell lines with defined pathogenic mutations, and secondly, by the establishment of a collection of patient tumor xenograft models with varied mtDNA status. Chapter two details the development of a new cytoplasmic hybrid (cybrid) generation method that simplifies the creation of transmitochondrial cybrids and enables transplantation of any mtDNA within any cell line. Utilizing this protocol, we developed isogenic melanoma lines with targeted mtDNA mutations. In chapter three, these melanoma cybrids are characterized in vivo, revealing that oxidative phosphorylation dysfunction does not block tumor growth. However, mtDNA mutations were found to significantly hinder spontaneous metastasis, largely due to impaired migration and invasion at metabolically restrictive conditions. Additionally, analysis of selective pressures exerted on the mitochondrial genomes in heteroplasmic cybrid cells demonstrated that tumor growth suppressed the pathogenic variant allele frequency (VAF) of mtDNA mutations. Lastly, chapter four analyzes renal cell carcinoma (RCC) patient-derived xenografts from the UT Southwestern Kidney Program, identifying RCC lines with either wildtype or pathogenic mtDNA mutations in Complex I subunits. RCC mtDNA mutations were validated through genetic and functional analyses. Additionally, it was found that RCC cancers with mtDNA mutations were associated with altered transcriptional statuses. Collectively, this thesis elucidates the relationship between mtDNA mutations and tumor development, employs innovative methods to examine mitochondrial function, and underscores the importance of mitochondrial genetics in oncology."],"dc:format":["application/pdf"],"dc:identifier":["https://hdl.handle.net/2152.5/10804","1596185292"],"dc:language":["en"],"dc:subject":["DNA, Mitochondrial","Melanoma","Mutation","Neoplasm Metastasis"],"dc:title":["Consequences of Pathogenic Mitochondrial DNA Mutations in Cancer Progression"],"dc:type":["Thesis","text"]},"updated_at":"2026-07-24T05:52:08Z"}