Objectives:Cancer treatment relies heavily on accurate diagnosis and effective monitoring of the disease.These processes often involve invasive procedures,such as colonoscopy,to detect malignant tissues,followed by mo...Objectives:Cancer treatment relies heavily on accurate diagnosis and effective monitoring of the disease.These processes often involve invasive procedures,such as colonoscopy,to detect malignant tissues,followed by molecular analyses to determine relevant biomarkers.This study aimed to evaluate the clinical performance of droplet digital PCR(ddPCR)for detecting Kirsten Rat Sarcoma Viral Proto-Oncogene(KRAS),Neuroblastoma RAS Viral Oncogene Homolog(NRAS),and B-Raf Murine Sarcoma Viral Oncogene Homolog B(BRAF)mutations in circulating tumor DNA(ctDNA)from colorectal cancer patients using liquid biopsy.Methods:ctDNA was isolated from colorectal cancer(CRC)patients(n=110)and analyzed for KRAS,BRAF,and NRAS mutations.The ctDNA obtained through liquid biopsy was analyzed using ddPCR,and the findings were compared with sequencing data from tumor DNA archived in formalin-fixed paraffin-embedded(FFPE)blocks.Results:For KRAS mutations,ddPCR achieved a sensitivity of 72.0%and a specificity of 71.4%.However,when pooling all target mutations(KRAS,NRAS and BRAF),the overall sensitivity and specificity were lower,at 48.3%and 51.1%,respectively.Conclusion:The results of this study indicate that the ddPCR analysis of ctDNA may provide complementary information for the molecular diagnosis of CRC patients.展开更多
Pancreatic ductal adenocarcinoma(PDAC)is an aggressive malignancy with a poor prognosis that is driven primarily by oncogenic KRAS mutations present in>90%of cases.KRAS mutations,particularly the G12D mutation whic...Pancreatic ductal adenocarcinoma(PDAC)is an aggressive malignancy with a poor prognosis that is driven primarily by oncogenic KRAS mutations present in>90%of cases.KRAS mutations,particularly the G12D mutation which dominates in PDAC,fuel tumor initiation,progression,and immune evasion,thereby contributing to therapy resistance.Nevertheless,KRAS has long been considered“undruggable”due to its structure.Recent advances have spurred transformative progress in direct KRAS inhibition.While FDAapproved mutation-specific and pan-KRAS inhibitors show limited efficacy in PDAC,emerging agents(MRTX1133 and RMC-9805)have demonstrated preclinical promise.However,resistance remains a critical hurdle and is driven by pathway reactivation,secondary mutations,and metabolic adaptations.Alternative strategies targeting upstream regulators(SHP2 and SOS1)aim to block KRAS activation and associated resistance mechanisms.Preclinical studies have also highlighted synergistic benefits of combining KRAS inhibitors with MEK,PI3K,or CDK4/6 inhibitors,which are now undergoing clinical evaluation.Immunotherapies,including KRAS-targeted vaccines and adoptive T-cell therapies,have further expanded the therapeutic landscape of enhancing KRAS-targeted therapies in PDAC.The molecular basis of KRAS-driven PDAC,current inhibitors,resistance mechanisms,and innovative strategies are discussed herein to address treatment barriers.Opportunities to improve clinical outcomes are underscored in this challenging malignancy by integrating insights from preclinical and clinical research.展开更多
基金funded by the Ministry of Health of the Czech Republic—conceptual development of research organization(MMCI,00209805)Czech Science Foundation(No.25-15990S)+1 种基金the project 7D241003 EUREKA EUROSTARS35897,project SALVAGE(P JAC,reg.No.CZ.02.01.01/00/22_008/0004644)—funded by the European Unionby the State Budget of the Czech Republic,and by the LRI project BBMRI.cz(Nos.LM2023033 and CZ.02.1.01/0.0/0.0/16_013/0001674.).
文摘Objectives:Cancer treatment relies heavily on accurate diagnosis and effective monitoring of the disease.These processes often involve invasive procedures,such as colonoscopy,to detect malignant tissues,followed by molecular analyses to determine relevant biomarkers.This study aimed to evaluate the clinical performance of droplet digital PCR(ddPCR)for detecting Kirsten Rat Sarcoma Viral Proto-Oncogene(KRAS),Neuroblastoma RAS Viral Oncogene Homolog(NRAS),and B-Raf Murine Sarcoma Viral Oncogene Homolog B(BRAF)mutations in circulating tumor DNA(ctDNA)from colorectal cancer patients using liquid biopsy.Methods:ctDNA was isolated from colorectal cancer(CRC)patients(n=110)and analyzed for KRAS,BRAF,and NRAS mutations.The ctDNA obtained through liquid biopsy was analyzed using ddPCR,and the findings were compared with sequencing data from tumor DNA archived in formalin-fixed paraffin-embedded(FFPE)blocks.Results:For KRAS mutations,ddPCR achieved a sensitivity of 72.0%and a specificity of 71.4%.However,when pooling all target mutations(KRAS,NRAS and BRAF),the overall sensitivity and specificity were lower,at 48.3%and 51.1%,respectively.Conclusion:The results of this study indicate that the ddPCR analysis of ctDNA may provide complementary information for the molecular diagnosis of CRC patients.
基金supported by the Tianshan Talents-Youth Science and Technology Innovation Talents Training Program of Xinjiang Autonomous Region(Grant No.2022TSYCCX0035)the Xinjiang Key Laboratory of Natural Medicines Active Components and Drug Release Technology(Grant No.XJDX1713)+2 种基金the“Fourteenth Five-Year Plan”Key Discipline Construction Project of Xinjiang Autonomous Region(2021)the Engineering Research Center of Xinjiang and Central Asian Medicine Resources,Ministry of Education(2023)the Xinjiang Key Laboratory of Biopharmaceuticals and Medical Devices(2023)。
文摘Pancreatic ductal adenocarcinoma(PDAC)is an aggressive malignancy with a poor prognosis that is driven primarily by oncogenic KRAS mutations present in>90%of cases.KRAS mutations,particularly the G12D mutation which dominates in PDAC,fuel tumor initiation,progression,and immune evasion,thereby contributing to therapy resistance.Nevertheless,KRAS has long been considered“undruggable”due to its structure.Recent advances have spurred transformative progress in direct KRAS inhibition.While FDAapproved mutation-specific and pan-KRAS inhibitors show limited efficacy in PDAC,emerging agents(MRTX1133 and RMC-9805)have demonstrated preclinical promise.However,resistance remains a critical hurdle and is driven by pathway reactivation,secondary mutations,and metabolic adaptations.Alternative strategies targeting upstream regulators(SHP2 and SOS1)aim to block KRAS activation and associated resistance mechanisms.Preclinical studies have also highlighted synergistic benefits of combining KRAS inhibitors with MEK,PI3K,or CDK4/6 inhibitors,which are now undergoing clinical evaluation.Immunotherapies,including KRAS-targeted vaccines and adoptive T-cell therapies,have further expanded the therapeutic landscape of enhancing KRAS-targeted therapies in PDAC.The molecular basis of KRAS-driven PDAC,current inhibitors,resistance mechanisms,and innovative strategies are discussed herein to address treatment barriers.Opportunities to improve clinical outcomes are underscored in this challenging malignancy by integrating insights from preclinical and clinical research.
