3 research outputs found
Establishment of patient-derived tumor organoids to functionally inform treatment decisions in metastatic colorectal cancer
Colorectal cancer; Personalized medicine; ResistanceC谩ncer colorrectal; Medicina personalizada; ResistenciaC脿ncer colorectal; Medicina personalitzada; Resist猫nciaBackground
Metastatic colorectal cancer (mCRC) patients tend to have modest benefits from molecularly driven therapeutics. Patient-derived tumor organoids (PDTOs) represent an unmatched model to elucidate tumor resistance to therapy, due to their high capacity to resemble tumor characteristics.
Materials and methods
We used viable tumor tissue from two cohorts of patients with mCRC, na茂ve or refractory to treatment, respectively, for generating PDTOs. The derived models were subjected to a 6-day drug screening assay (DSA) with a comprehensive pipeline of chemotherapy and targeted drugs against almost all the actionable mCRC molecular drivers. For the second cohort DSA data were matched with those from PDTO genotyping.
Results
A total of 40 PDTOs included in the two cohorts were derived from mCRC primary tumors or metastases. The first cohort included 31 PDTOs derived from patients treated in front line. For this cohort, DSA results were matched with patient responses. Moreover, RAS/BRAF mutational status was matched with DSA cetuximab response. Ten out of 12 (83.3%) RAS wild-type PDTOs responded to cetuximab, while all the mutant PDTOs, 8 out of 8 (100%), were resistant. For the second cohort (chemorefractory patients), we used part of tumor tissue for genotyping. Four out of nine DSA/genotyping data resulted applicable in the clinic. Two RAS-mutant mCRC patients have been treated with FOLFOX鈥揵evacizumab and mitomycin鈥揷apecitabine in third line, respectively, based on DSA results, obtaining disease control. One patient was treated with nivolumab鈥搒econd mitochondrial-derived activator of caspases mimetic (phase I trial) due to high tumor mutational burden at genotyping, experiencing stable disease. In one case, the presence of BRCA2 mutation correlated with DSA sensitivity to olaparib; however, the patient could not receive the therapy.
Conclusions
Using CRC as a model, we have designed and validated a clinically applicable methodology to potentially inform clinical decisions with functional data. Undoubtedly, further larger analyses are needed to improve methodology success rates and propose suitable treatment strategies for mCRC patients.This Translational Research Fellowship Project was supported by the ESMO with the aid of a grant from Amgen, by the Accelerator (ACRCelerator) [grant number A26825] and Ayuda a m茅dicos j贸venes investigadores from Fundacion Cient铆fica鈥擜sociacion Espa帽ola Contra el Cancer (FC-AECC)/Associazione Italiana per la Ricerca sul Cancro (AIRC)/Cancer Research United Kingdom (CRUK) and by Familia Armangu茅. Any views, opinions, findings, conclusions or recommendations expressed in this material are those solely of the author(s) and do not necessarily reflect those of ESMO or Amgen. We thank Regione Campania (I-Cure Research Project) [grant number: Cup 21C17000030007], ESMO Translational Research Fellowship Program
Mecanismos de resistencia a terapias dirigidas contra BRAF(V600E) y estudio de nuevas combinaciones de f谩rmacos en c谩ncer de colon
Programa de Doctorat en Biomedicina / Tesi realitzada a l'Institut d'Oncologia Vall d'Hebron (VHIO)[spa] Alrededor del 10% de los pacientes con c谩ncer colorrectal metast谩tico (mCRC) presentan la mutaci贸n BRAF(V600E), mutaci贸n que est谩 asociada a una peor respuesta al tratamiento est谩ndar con agentes quimioterap茅uticos y a una menor supervivencia. Recientemente se ha aprobado el uso de la combinaci贸n encorafenib + cetuximab para el tratamiento del mCRC BRAF(V600E). Desafortunadamente, s贸lo un peque帽o porcentaje de los pacientes responden a este tratamiento y una importante fracci贸n de 茅stos recaen al poco tiempo, lo que hace que sea muy importante identificar los mecanismos asociados a la adquisici贸n de resistencia a los inhibidores de BRAF (BRAFi) y la identificaci贸n de nuevas vulnerabilidades. En este proyecto, hemos descrito que en respuesta a terapias combinadas con BRAFi en CRC se da una inducci贸n del programa hip贸xico y angiog茅nico, la activaci贸n de los fibroblastos asociados al c谩ncer (CAFs) y una transici贸n a un fenotipo extremadamente mucinoso. Donde, la calidad de la ramificaci贸n de la estructura de las mucinas es necesaria para la adquisici贸n de resistencia a las terapias BRAFi.
Adem谩s, hemos observado un aumento de la expresi贸n de MMP7, el cual promueve y mejora la liberaci贸n y accesibilidad de VEGFA bioactivo, lo que induce la angiog茅nesis tras el tratamiento en modelos precl铆nicos PDX y l铆neas celulares tratados con encorafenib + cetuximab + binimetinib. Estos acontecimientos generan una vulnerabilidad que nos dio la oportunidad de dise帽ar nuevas combinaciones racionales y efectivas a帽adiendo un inhibidor de VEGFA (VEGFAi) al tratamiento. Esta nueva combinaci贸n con el anticuerpo anti-angiog茅nico bevacizumab retras贸 el tiempo de progresi贸n de las terapias BRAFi y nos ha permitido establecer un ensayo cl铆nico nuevo para validar este nuevo r茅gimen terap茅utico en pacientes de CRC con tumores BRAF(V600E).
Paralelamente, hemos descrito que en pacientes de CRC la hipermetilaci贸n del promotor del gen BRCA1 puede reprimir su expresi贸n, liderando as铆 una deficiencia en la reparaci贸n por recombinaci贸n hom贸loga (HRD) y una sensibilidad a los inhibidores de PARP (PARPi). Adem谩s, hemos descrito de forma detallada un caso de CRC en el que el tratamiento y la progresi贸n a los BRAFi da lugar a la selecci贸n de clones con una reparaci贸n hom贸loga funcional (HRP), los cuales proven铆an de una enfermedad que inicialmente presentaba HRD. De este caso, hemos extra铆do una firma de metilaci贸n que nos permitir铆a identificar casos con HRD de diferentes tipos tumorales. Esto es especialmente evidente en tumores donde la HRD es una alteraci贸n frecuente, como es el caso del adenocarcinoma de pulm贸n o el c谩ncer de ovario. Los datos obtenidos de este paciente apoyan el concepto de tratar a pacientes de CRC con PARPi mientras presenten una HRD. Son necesarios m谩s estudios para refinar y validar el potencial de nuestra firma epigen茅tica como marcador de selecci贸n de pacientes HRD sensibles a PARPi.[eng] Around 10% of patients with metastatic colorectal cancer (mCRC) present a BRAF(V600E) mutation, which is associated with a poor response to standard chemotherapy and shorter overall survival. The combination encorafenib + cetuximab has been recently approved for the treatment of mCRC BRAF(V600E). Unfortunately, only a small percentage of them respond to this treatment and a large fraction of patients relapse shortly, so it has become very important to identify the mechanisms associated to the acquired resistance to BRAF inhibitors (BRAFi) and the identification of new vulnerabilities. In this project, we described a hypoxic and angiogenic program induction, the activation of tumour associated fibroblasts (CAFs) and the transition to an extreme mucinous differentiation phenotype as a response to BRAFi- combined therapies in CRC. This drastic tumour tissue remodelling leads to a non-genetic drug resistance. In particular the quality of the branched structure in mucins is required for the acquisition of resistance to BRAFi therapies.
Furthermore, we observed an increase in MMP7 expression that promoted an enhanced release of bioactive VEGFA accessible to induce angiogenesis upon treatment of preclinical PDX and cell line models with encorafenib + cetuximab + binimetinib. These events became a vulnerability that gave us the opportunity to design new rational and effective combinations adding a VEGFA inhibitor (VEGFAi) to the treatment. This new combination with the anti- angiogenic antibody bevacizumab delayed the time to progression to BRAFi therapy and led us to set a new clinical trial to validate such therapeutic regime in CRC patients with BRAF(V600E) tumours.
In parallel, we describe that in CRC patients the hypermethylation of BRCA1 gene promoter can repress its expression leading to Homologous Repair Deficiency (HRD) and sensitivity to PARP inhibitors (PARPi). Here we also report in detail a CRC case in which treatment and progression to BRAF(V600E) inhibitory therapy selected Homologous Repair Proficient (HRP) clones from an initial disease presenting HRD. From this case, we extracted a methylation signature that could identify HRD cases suffering of different tumour types. This is particularly evident in tumours where HRD has been described as a frequent alteration, such as lung adenocarcinomas or ovarian cancer. Our real patient data supports the concept of treating CRC patients with PARP inhibitors as far as they present HRD. Future studies will be required to refine and validate the potential of our epigenetic signature as biomarker for selecting HRD patients sensitive to PARP inhibitor
Mecanismos de resistencia a terapias dirigidas contra BRAF(V600E) y estudio de nuevas combinaciones de f谩rmacos en c谩ncer de colon
[spa] Alrededor del 10% de los pacientes con c谩ncer colorrectal metast谩tico (mCRC) presentan la mutaci贸n BRAF(V600E), mutaci贸n que est谩 asociada a una peor respuesta al tratamiento est谩ndar con agentes quimioterap茅uticos y a una menor supervivencia. Recientemente se ha aprobado el uso de la combinaci贸n encorafenib + cetuximab para el tratamiento del mCRC BRAF(V600E). Desafortunadamente, s贸lo un peque帽o porcentaje de los pacientes responden a este tratamiento y una importante fracci贸n de 茅stos recaen al poco tiempo, lo que hace que sea muy importante identificar los mecanismos asociados a la adquisici贸n de resistencia a los inhibidores de BRAF (BRAFi) y la identificaci贸n de nuevas vulnerabilidades. En este proyecto, hemos descrito que en respuesta a terapias combinadas con BRAFi en CRC se da una inducci贸n del programa hip贸xico y angiog茅nico, la activaci贸n de los fibroblastos asociados al c谩ncer (CAFs) y una transici贸n a un fenotipo extremadamente mucinoso. Donde, la calidad de la ramificaci贸n de la estructura de las mucinas es necesaria para la adquisici贸n de resistencia a las terapias BRAFi. Adem谩s, hemos observado un aumento de la expresi贸n de MMP7, el cual promueve y mejora la liberaci贸n y accesibilidad de VEGFA bioactivo, lo que induce la angiog茅nesis tras el tratamiento en modelos precl铆nicos PDX y l铆neas celulares tratados con encorafenib + cetuximab + binimetinib. Estos acontecimientos generan una vulnerabilidad que nos dio la oportunidad de dise帽ar nuevas combinaciones racionales y efectivas a帽adiendo un inhibidor de VEGFA (VEGFAi) al tratamiento. Esta nueva combinaci贸n con el anticuerpo anti-angiog茅nico bevacizumab retras贸 el tiempo de progresi贸n de las terapias BRAFi y nos ha permitido establecer un ensayo cl铆nico nuevo para validar este nuevo r茅gimen terap茅utico en pacientes de CRC con tumores BRAF(V600E). Paralelamente, hemos descrito que en pacientes de CRC la hipermetilaci贸n del promotor del gen BRCA1 puede reprimir su expresi贸n, liderando as铆 una deficiencia en la reparaci贸n por recombinaci贸n hom贸loga (HRD) y una sensibilidad a los inhibidores de PARP (PARPi). Adem谩s, hemos descrito de forma detallada un caso de CRC en el que el tratamiento y la progresi贸n a los BRAFi da lugar a la selecci贸n de clones con una reparaci贸n hom贸loga funcional (HRP), los cuales proven铆an de una enfermedad que inicialmente presentaba HRD. De este caso, hemos extra铆do una firma de metilaci贸n que nos permitir铆a identificar casos con HRD de diferentes tipos tumorales. Esto es especialmente evidente en tumores donde la HRD es una alteraci贸n frecuente, como es el caso del adenocarcinoma de pulm贸n o el c谩ncer de ovario. Los datos obtenidos de este paciente apoyan el concepto de tratar a pacientes de CRC con PARPi mientras presenten una HRD. Son necesarios m谩s estudios para refinar y validar el potencial de nuestra firma epigen茅tica como marcador de selecci贸n de pacientes HRD sensibles a PARPi.[eng] Around 10% of patients with metastatic colorectal cancer (mCRC) present a BRAF(V600E) mutation, which is associated with a poor response to standard chemotherapy and shorter overall survival. The combination encorafenib + cetuximab has been recently approved for the treatment of mCRC BRAF(V600E). Unfortunately, only a small percentage of them respond to this treatment and a large fraction of patients relapse shortly, so it has become very important to identify the mechanisms associated to the acquired resistance to BRAF inhibitors (BRAFi) and the identification of new vulnerabilities. In this project, we described a hypoxic and angiogenic program induction, the activation of tumour associated fibroblasts (CAFs) and the transition to an extreme mucinous differentiation phenotype as a response to BRAFi- combined therapies in CRC. This drastic tumour tissue remodelling leads to a non-genetic drug resistance. In particular the quality of the branched structure in mucins is required for the acquisition of resistance to BRAFi therapies. Furthermore, we observed an increase in MMP7 expression that promoted an enhanced release of bioactive VEGFA accessible to induce angiogenesis upon treatment of preclinical PDX and cell line models with encorafenib + cetuximab + binimetinib. These events became a vulnerability that gave us the opportunity to design new rational and effective combinations adding a VEGFA inhibitor (VEGFAi) to the treatment. This new combination with the anti- angiogenic antibody bevacizumab delayed the time to progression to BRAFi therapy and led us to set a new clinical trial to validate such therapeutic regime in CRC patients with BRAF(V600E) tumours. In parallel, we describe that in CRC patients the hypermethylation of BRCA1 gene promoter can repress its expression leading to Homologous Repair Deficiency (HRD) and sensitivity to PARP inhibitors (PARPi). Here we also report in detail a CRC case in which treatment and progression to BRAF(V600E) inhibitory therapy selected Homologous Repair Proficient (HRP) clones from an initial disease presenting HRD. From this case, we extracted a methylation signature that could identify HRD cases suffering of different tumour types. This is particularly evident in tumours where HRD has been described as a frequent alteration, such as lung adenocarcinomas or ovarian cancer. Our real patient data supports the concept of treating CRC patients with PARP inhibitors as far as they present HRD. Future studies will be required to refine and validate the potential of our epigenetic signature as biomarker for selecting HRD patients sensitive to PARP inhibitor