{"id":{"repo_id":"milano","oai_identifier":"oai:air.unimi.it:2434/1197878"},"canonical_url":"https://search.dev.ndltd.org/etd/milano/oai:air.unimi.it:2434/1197878","repository":{"repo_id":"milano","name":"Università degli Studi di Milano","base_url":"https://air.unimi.it/oai/request"},"display":{"title":"TARGETING CANCER METABOLISM WITH FASTING-MIMICKING DIET¿BASED LOW-TOXIC COMBINATION THERAPIES: OVERCOMING RESISTANCE THROUGH METABOLIC SYNERGY","abstract":"Aging represents the highest risk factor for cancer, defined by molecular hallmarks such as genomic instability, telomere attrition, deregulated nutrient sensing, and chronic inflammation promoting malignant transformation. Among age-associated cancers, non-small cell lung cancer (NSCLC) is the most prevalent and lethal, with KRAS mutations occurring in up to 25–30% of lung adenocarcinomas. Tumors harbouring this mutated oncogene are particularly resistant to targeted therapies, highlighting the need for novel targeted low-toxicity strategies. Nutrient restriction interventions, such as fasting and fasting-mimicking diets (FMD), modulate conserved growth pathways (IGF-1/AKT/mTOR, AMPK) and enhance cellular stress resistance, offering a promising approach to cancer therapy. FMD, if combined with treatments like pharmacological doses of ascorbate could effectively target NSCLC rewired metabolism that sustains NSCLC progression. In this thesis, I investigated the combinatorial effects of FMD and pharmacological doses of vitamin C in KRAS-mutant NSCLC. In vitro, short-term starvation (STS) synergized with vitamin C to induce dramatic cell death in KRAS-mutant and wild- type NSCLC cell lines, while protecting normal cells. In vivo, cycles of FMD combined with high-dose vitamin C significantly delayed tumor growth in both immunodeficient (NSG) and immunocompetent (C57BL/6) mouse models, without inducing major toxicity. However, as previously demonstrated in our studies, cancer cells resist FMD through upregulation of starvation escape pathways. Here, mechanistically, RNA sequencing (RNA-seq) analysis revealed that resistant subpopulations upregulated MAPK signaling as an acute adaptive response, while chronic exposure drove additional activation of PI3K-AKT pathways to resist FMD and vitamin C treatment. Pharmacological blockade of EGFR/ERK (Afatinib, Ulixertinib) or PI3K (Pictilisib), particularly in dual ERK/PI3K inhibition, synergized with FMD and vitamin C to overcome resistance, delay tumor progression, and induce tumor regression in vivo. Collectively, these findings identify the FMD–vitamin C axis as a safe and effective metabolic therapy for NSCLC and highlight MAPK and PI3K signaling as key resistance drivers. Rational combination of metabolic interventions with targeted inhibitors provides a promising framework for personalized strategies in KRAS- driven lung cancer.","abstract_html":"Aging represents the highest risk factor for cancer, defined by molecular hallmarks such as genomic instability, telomere attrition, deregulated nutrient sensing, and chronic inflammation promoting malignant transformation. Among age-associated cancers, non-small cell lung cancer (NSCLC) is the most prevalent and lethal, with KRAS mutations occurring in up to 25–30% of lung adenocarcinomas. Tumors harbouring this mutated oncogene are particularly resistant to targeted therapies, highlighting the need for novel targeted low-toxicity strategies. Nutrient restriction interventions, such as fasting and fasting-mimicking diets (FMD), modulate conserved growth pathways (IGF-1/AKT/mTOR, AMPK) and enhance cellular stress resistance, offering a promising approach to cancer therapy. FMD, if combined with treatments like pharmacological doses of ascorbate could effectively target NSCLC rewired metabolism that sustains NSCLC progression. In this thesis, I investigated the combinatorial effects of FMD and pharmacological doses of vitamin C in KRAS-mutant NSCLC. In vitro, short-term starvation (STS) synergized with vitamin C to induce dramatic cell death in KRAS-mutant and wild- type NSCLC cell lines, while protecting normal cells. In vivo, cycles of FMD combined with high-dose vitamin C significantly delayed tumor growth in both immunodeficient (NSG) and immunocompetent (C57BL/6) mouse models, without inducing major toxicity. However, as previously demonstrated in our studies, cancer cells resist FMD through upregulation of starvation escape pathways. Here, mechanistically, RNA sequencing (RNA-seq) analysis revealed that resistant subpopulations upregulated MAPK signaling as an acute adaptive response, while chronic exposure drove additional activation of PI3K-AKT pathways to resist FMD and vitamin C treatment. Pharmacological blockade of EGFR/ERK (Afatinib, Ulixertinib) or PI3K (Pictilisib), particularly in dual ERK/PI3K inhibition, synergized with FMD and vitamin C to overcome resistance, delay tumor progression, and induce tumor regression in vivo. Collectively, these findings identify the FMD–vitamin C axis as a safe and effective metabolic therapy for NSCLC and highlight MAPK and PI3K signaling as key resistance drivers. Rational combination of metabolic interventions with targeted inhibitors provides a promising framework for personalized strategies in KRAS- driven lung cancer.","abstract_has_math":false,"creators":["SHMAHALA, ANASTASIYA"],"institution":"Università degli Studi di Milano","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["tutor: V. D. Longo ; co-tutor: V. Costanzo ; internal advisor: S. Casola ; external advisor: M. Vinciguerra","Casola","Stefano; Manlio","Vinciguerra","A. Shmahala","COSTANZO, VINCENZO"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-12-17","date_published":"2025-12-17","updated_at":"2026-07-27T20:19:08Z","subjects":["oncology","nutrition","fasting","diet","resistance","oxidative stre","lung cancer","KRAS oncogene","ascorbate","metabolism","Settore MEDS-02/A - Patologia generale"],"languages":["eng"],"rights":["info:eu-repo/semantics/embargoedAccess","license:Creative commons","license uri:http://creativecommons.org/licenses/by-sa/4.0/"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/2434/1197878","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["tutor: V. D. Longo ; co-tutor: V. Costanzo ; internal advisor: S. Casola ; external advisor: M. Vinciguerra","Casola","Stefano; Manlio","Vinciguerra","A. Shmahala","COSTANZO, VINCENZO"]},{"key":"dc:creator","label":"Author","values":["SHMAHALA, ANASTASIYA"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2025-12-17"]},{"key":"dc:publisher","label":"Institution","values":["Università degli Studi di Milano","place:IFOM ETS - The AIRC Institute of Molecular Oncology"]},{"key":"dc:relation","label":"Dc Relation","values":["numberofpages:112","alleditors:Casola, Stefano; Manlio, Vinciguerra"]},{"key":"dc:type","label":"Dc Type","values":["info:eu-repo/semantics/doctoralThesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["oncology","nutrition","fasting","diet","resistance","oxidative stre","lung cancer","KRAS oncogene","ascorbate","metabolism","Settore MEDS-02/A - Patologia generale"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["info:eu-repo/semantics/embargoedAccess","license:Creative commons","license uri:http://creativecommons.org/licenses/by-sa/4.0/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://hdl.handle.net/2434/1197878"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Aging represents the highest risk factor for cancer, defined by molecular hallmarks such as genomic instability, telomere attrition, deregulated nutrient sensing, and chronic inflammation promoting malignant transformation. Among age-associated cancers, non-small cell lung cancer (NSCLC) is the most prevalent and lethal, with KRAS mutations occurring in up to 25–30% of lung adenocarcinomas. Tumors harbouring this mutated oncogene are particularly resistant to targeted therapies, highlighting the need for novel targeted low-toxicity strategies. Nutrient restriction interventions, such as fasting and fasting-mimicking diets (FMD), modulate conserved growth pathways (IGF-1/AKT/mTOR, AMPK) and enhance cellular stress resistance, offering a promising approach to cancer therapy. FMD, if combined with treatments like pharmacological doses of ascorbate could effectively target NSCLC rewired metabolism that sustains NSCLC progression. In this thesis, I investigated the combinatorial effects of FMD and pharmacological doses of vitamin C in KRAS-mutant NSCLC. In vitro, short-term starvation (STS) synergized with vitamin C to induce dramatic cell death in KRAS-mutant and wild- type NSCLC cell lines, while protecting normal cells. In vivo, cycles of FMD combined with high-dose vitamin C significantly delayed tumor growth in both immunodeficient (NSG) and immunocompetent (C57BL/6) mouse models, without inducing major toxicity. However, as previously demonstrated in our studies, cancer cells resist FMD through upregulation of starvation escape pathways. Here, mechanistically, RNA sequencing (RNA-seq) analysis revealed that resistant subpopulations upregulated MAPK signaling as an acute adaptive response, while chronic exposure drove additional activation of PI3K-AKT pathways to resist FMD and vitamin C treatment. Pharmacological blockade of EGFR/ERK (Afatinib, Ulixertinib) or PI3K (Pictilisib), particularly in dual ERK/PI3K inhibition, synergized with FMD and vitamin C to overcome resistance, delay tumor progression, and induce tumor regression in vivo. Collectively, these findings identify the FMD–vitamin C axis as a safe and effective metabolic therapy for NSCLC and highlight MAPK and PI3K signaling as key resistance drivers. Rational combination of metabolic interventions with targeted inhibitors provides a promising framework for personalized strategies in KRAS- driven lung cancer."]},{"key":"dc:title","label":"Title","values":["TARGETING CANCER METABOLISM WITH FASTING-MIMICKING DIET¿BASED LOW-TOXIC COMBINATION THERAPIES: OVERCOMING RESISTANCE THROUGH METABOLIC SYNERGY"]}]}],"canonical_facts":{"dc:contributor":["tutor: V. D. Longo ; co-tutor: V. Costanzo ; internal advisor: S. Casola ; external advisor: M. Vinciguerra","Casola","Stefano; Manlio","Vinciguerra","A. Shmahala","COSTANZO, VINCENZO"],"dc:creator":["SHMAHALA, ANASTASIYA"],"dc:date":["2025-12-17"],"dc:description":["Aging represents the highest risk factor for cancer, defined by molecular hallmarks such as genomic instability, telomere attrition, deregulated nutrient sensing, and chronic inflammation promoting malignant transformation. Among age-associated cancers, non-small cell lung cancer (NSCLC) is the most prevalent and lethal, with KRAS mutations occurring in up to 25–30% of lung adenocarcinomas. Tumors harbouring this mutated oncogene are particularly resistant to targeted therapies, highlighting the need for novel targeted low-toxicity strategies. Nutrient restriction interventions, such as fasting and fasting-mimicking diets (FMD), modulate conserved growth pathways (IGF-1/AKT/mTOR, AMPK) and enhance cellular stress resistance, offering a promising approach to cancer therapy. FMD, if combined with treatments like pharmacological doses of ascorbate could effectively target NSCLC rewired metabolism that sustains NSCLC progression. In this thesis, I investigated the combinatorial effects of FMD and pharmacological doses of vitamin C in KRAS-mutant NSCLC. In vitro, short-term starvation (STS) synergized with vitamin C to induce dramatic cell death in KRAS-mutant and wild- type NSCLC cell lines, while protecting normal cells. In vivo, cycles of FMD combined with high-dose vitamin C significantly delayed tumor growth in both immunodeficient (NSG) and immunocompetent (C57BL/6) mouse models, without inducing major toxicity. However, as previously demonstrated in our studies, cancer cells resist FMD through upregulation of starvation escape pathways. Here, mechanistically, RNA sequencing (RNA-seq) analysis revealed that resistant subpopulations upregulated MAPK signaling as an acute adaptive response, while chronic exposure drove additional activation of PI3K-AKT pathways to resist FMD and vitamin C treatment. Pharmacological blockade of EGFR/ERK (Afatinib, Ulixertinib) or PI3K (Pictilisib), particularly in dual ERK/PI3K inhibition, synergized with FMD and vitamin C to overcome resistance, delay tumor progression, and induce tumor regression in vivo. Collectively, these findings identify the FMD–vitamin C axis as a safe and effective metabolic therapy for NSCLC and highlight MAPK and PI3K signaling as key resistance drivers. Rational combination of metabolic interventions with targeted inhibitors provides a promising framework for personalized strategies in KRAS- driven lung cancer."],"dc:identifier":["https://hdl.handle.net/2434/1197878"],"dc:language":["eng"],"dc:publisher":["Università degli Studi di Milano","place:IFOM ETS - The AIRC Institute of Molecular Oncology"],"dc:relation":["numberofpages:112","alleditors:Casola, Stefano; Manlio, Vinciguerra"],"dc:rights":["info:eu-repo/semantics/embargoedAccess","license:Creative commons","license uri:http://creativecommons.org/licenses/by-sa/4.0/"],"dc:subject":["oncology","nutrition","fasting","diet","resistance","oxidative stre","lung cancer","KRAS oncogene","ascorbate","metabolism","Settore MEDS-02/A - Patologia generale"],"dc:title":["TARGETING CANCER METABOLISM WITH FASTING-MIMICKING DIET¿BASED LOW-TOXIC COMBINATION THERAPIES: OVERCOMING RESISTANCE THROUGH METABOLIC SYNERGY"],"dc:type":["info:eu-repo/semantics/doctoralThesis"]},"updated_at":"2026-07-27T20:19:08Z"}