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1.
Ann Oncol ; 34(9): 813-825, 2023 09.
Article de Anglais | MEDLINE | ID: mdl-37330052

RÉSUMÉ

BACKGROUND: The isolation of cell-free DNA (cfDNA) from the bloodstream can be used to detect and analyze somatic alterations in circulating tumor DNA (ctDNA), and multiple cfDNA-targeted sequencing panels are now commercially available for Food and Drug Administration (FDA)-approved biomarker indications to guide treatment. More recently, cfDNA fragmentation patterns have emerged as a tool to infer epigenomic and transcriptomic information. However, most of these analyses used whole-genome sequencing, which is insufficient to identify FDA-approved biomarker indications in a cost-effective manner. PATIENTS AND METHODS: We used machine learning models of fragmentation patterns at the first coding exon in standard targeted cancer gene cfDNA sequencing panels to distinguish between cancer and non-cancer patients, as well as the specific tumor type and subtype. We assessed this approach in two independent cohorts: a published cohort from GRAIL (breast, lung, and prostate cancers, non-cancer, n = 198) and an institutional cohort from the University of Wisconsin (UW; breast, lung, prostate, bladder cancers, n = 320). Each cohort was split 70%/30% into training and validation sets. RESULTS: In the UW cohort, training cross-validated accuracy was 82.1%, and accuracy in the independent validation cohort was 86.6% despite a median ctDNA fraction of only 0.06. In the GRAIL cohort, to assess how this approach performs in very low ctDNA fractions, training and independent validation were split based on ctDNA fraction. Training cross-validated accuracy was 80.6%, and accuracy in the independent validation cohort was 76.3%. In the validation cohort where the ctDNA fractions were all <0.05 and as low as 0.0003, the cancer versus non-cancer area under the curve was 0.99. CONCLUSIONS: To our knowledge, this is the first study to demonstrate that sequencing from targeted cfDNA panels can be utilized to analyze fragmentation patterns to classify cancer types, dramatically expanding the potential capabilities of existing clinically used panels at minimal additional cost.


Sujet(s)
Acides nucléiques acellulaires , ADN tumoral circulant , Tumeurs de la prostate , Mâle , Humains , ADN tumoral circulant/génétique , Mutation , Tumeurs de la prostate/génétique , Acides nucléiques acellulaires/génétique , Analyse de profil d'expression de gènes , Marqueurs biologiques tumoraux/génétique
2.
Ann Oncol ; 29(2): 352-360, 2018 02 01.
Article de Anglais | MEDLINE | ID: mdl-29069303

RÉSUMÉ

Background: Genomic aberrations have been identified in metastatic castration-resistant prostate cancer (mCRPC), but molecular predictors of resistance to abiraterone acetate/prednisone (AA/P) treatment are not known. Patients and methods: In a prospective clinical trial, mCRPC patients underwent whole-exome sequencing (n = 82) and RNA sequencing (n = 75) of metastatic biopsies before initiating AA/P with the objective of identifying genomic alterations associated with resistance to AA/P. Primary resistance was determined at 12 weeks of treatment using criteria for progression that included serum prostate-specific antigen measurement, bone and computerized tomography imaging and symptom assessments. Acquired resistance was determined using the end point of time to treatment change (TTTC), defined as time from enrollment until change in treatment from progressive disease. Associations of genomic and transcriptomic alterations with primary resistance were determined using logistic regression, Fisher's exact test, single and multivariate analyses. Cox regression models were utilized for determining association of genomic and transcriptomic alterations with TTTC. Results: At 12 weeks, 32 patients in the cohort had progressed (nonresponders). Median study follow-up was 32.1 months by which time 58 patients had switched treatments due to progression. Median TTTC was 10.1 months (interquartile range: 4.4-24.1). Genes in the Wnt/ß-catenin pathway were more frequently mutated and negative regulators of Wnt/ß-catenin signaling were more frequently deleted or displayed reduced mRNA expression in nonresponders. Additionally, mRNA expression of cell cycle regulatory genes was increased in nonresponders. In multivariate models, increased cell cycle proliferation scores (≥ 50) were associated with shorter TTTC (hazard ratio = 2.11, 95% confidence interval: 1.17-3.80; P = 0.01). Conclusions: Wnt/ß-catenin pathway activation and increased cell cycle progression scores can serve as molecular markers for predicting resistance to AA/P therapy.


Sujet(s)
Acétate d'abiratérone/administration et posologie , Résistance aux médicaments antinéoplasiques/génétique , Prednisone/administration et posologie , Tumeurs prostatiques résistantes à la castration/génétique , Voie de signalisation Wnt/génétique , Sujet âgé , Protocoles de polychimiothérapie antinéoplasique/usage thérapeutique , Cycle cellulaire , Prolifération cellulaire , Étude d'association pangénomique , Humains , Mâle , Adulte d'âge moyen , Métastase tumorale/traitement médicamenteux , Métastase tumorale/génétique , Études prospectives , Tumeurs prostatiques résistantes à la castration/traitement médicamenteux
3.
Prostate Cancer Prostatic Dis ; 19(3): 231-41, 2016 09.
Article de Anglais | MEDLINE | ID: mdl-27184811

RÉSUMÉ

While there are myriad mechanisms of primary and acquired resistance to conventional and next-generation hormonal therapies in prostate cancer, the potential role of androgen receptor splice variants (AR-Vs) has recently gained momentum. AR-Vs are abnormally truncated isoforms of the androgen receptor (AR) protein that lack the COOH-terminal domain but retain the NH2-terminal domain and DNA-binding domain and are thus constitutively active even in the absence of ligands. Although multiple preclinical studies have previously implicated AR-Vs in the development of castration resistance as well as resistance to abiraterone and enzalutamide, recent technological advances have made it possible to reliably detect and quantify AR-Vs from human clinical tumor specimens including blood samples. Initial clinical studies have now shown that certain AR-Vs, in particular AR-V7, may be associated with resistance to abiraterone and enzalutamide but not taxane chemotherapies when detected in circulating tumor cells. Efforts are now underway to clinically validate AR-V7 as a relevant treatment-selection biomarker in the context of other key genomic aberrations in men with metastatic castration-resistant prostate cancer. Additional efforts are underway to therapeutically target both AR and AR-Vs either directly or indirectly. Whether AR-Vs represent drivers of castration-resistant prostate cancer, or whether they are simply passenger events associated with aggressive disease or clonal heterogeneity, will ultimately be answered only through these types of clinical trials.


Sujet(s)
Transformation cellulaire néoplasique/génétique , Variation génétique , Tumeurs de la prostate/génétique , Tumeurs de la prostate/métabolisme , Récepteurs aux androgènes/génétique , Récepteurs aux androgènes/métabolisme , Épissage alternatif , Antagonistes du récepteur des androgènes/usage thérapeutique , Androgènes/métabolisme , Animaux , Antinéoplasiques hormonaux/usage thérapeutique , Marqueurs biologiques tumoraux , Transformation cellulaire néoplasique/métabolisme , Essais cliniques comme sujet , Évaluation préclinique de médicament , Résistance aux médicaments antinéoplasiques/effets des médicaments et des substances chimiques , Épithélium/métabolisme , Épithélium/anatomopathologie , Régulation de l'expression des gènes tumoraux , Humains , Mâle , Thérapie moléculaire ciblée , Tumeurs de la prostate/traitement médicamenteux , Tumeurs de la prostate/anatomopathologie , Liaison aux protéines , Motifs et domaines d'intéraction protéique , Multimérisation de protéines , Récepteurs aux androgènes/composition chimique , Recherche , Transduction du signal/effets des médicaments et des substances chimiques , Transcription génétique , Résultat thérapeutique
4.
Oncogene ; 31(45): 4759-67, 2012 Nov 08.
Article de Anglais | MEDLINE | ID: mdl-22266865

RÉSUMÉ

Reactivation of the androgen receptor (AR) during androgen depletion therapy (ADT) underlies castration-resistant prostate cancer (CRPCa). Alternative splicing of the AR gene and synthesis of constitutively active COOH-terminally truncated AR variants lacking the AR ligand-binding domain has emerged as an important mechanism of ADT resistance in CRPCa. In a previous study, we demonstrated that altered AR splicing in CRPCa 22Rv1 cells was linked to a 35-kb intragenic tandem duplication of AR exon 3 and flanking sequences. In this study, we demonstrate that complex patterns of AR gene copy number imbalances occur in PCa cell lines, xenografts and clinical specimens. To investigate whether these copy number imbalances reflect AR gene rearrangements that could be linked to splicing disruptions, we carried out a detailed analysis of AR gene structure in the LuCaP 86.2 and CWR-R1 models of CRPCa. By deletion-spanning PCR, we discovered a 8579-bp deletion of AR exons 5, 6 and 7 in the LuCaP 86.2 xenograft, which provides a rational explanation for synthesis of the truncated AR v567es AR variant in this model. Similarly, targeted resequencing of the AR gene in CWR-R1 cells led to the discovery of a 48-kb deletion in AR intron 1. This intragenic deletion marked a specific CWR-R1 cell population with enhanced expression of the truncated AR-V7/AR3 variant, a high level of androgen-independent AR transcriptional activity and rapid androgen independent growth. Together, these data demonstrate that structural alterations in the AR gene are linked to stable gain-of-function splicing alterations in CRPCa.


Sujet(s)
Épissage alternatif , Délétion de gène , Régulation de l'expression des gènes tumoraux , Tumeurs de la prostate/génétique , Récepteurs aux androgènes/génétique , Animaux , Lignée cellulaire tumorale , Variations de nombre de copies de segment d'ADN , Modèles animaux de maladie humaine , Exons , Ordre des gènes , Humains , Introns , Mâle , Souris , Orchidectomie , Tumeurs de la prostate/métabolisme , Stabilité de l'ARN , Récepteurs aux androgènes/métabolisme , Transplantation hétérologue
5.
J Biol Chem ; 275(48): 37604-11, 2000 Dec 01.
Article de Anglais | MEDLINE | ID: mdl-10978326

RÉSUMÉ

The SRC gene encodes the proto-oncogene pp60(c-)(src), a tyrosine kinase implicated in numerous signal transduction pathways. In addition, the SRC gene is differentially expressed, developmentally regulated, and frequently overexpressed in human neoplasia. However, the mechanisms regulating its expression have not been completely explored. Here we describe the isolation of a new distal SRC promoter and associated exon, designated 1alpha, which we mapped to a position 1.0 kilobase upstream of the previously described SRC1A housekeeping promoter. Differential use of these promoters and their associated exons coupled with subsequent splicing to a common downstream exon results in c-Src transcripts with different 5' ends but identical coding regions. Promoter analysis following transient transfections into HepG2 cells mapped the minimal 1alpha promoter to a region 145 bp upstream of the major transcription start site. This region contained a consensus binding site for hepatic nuclear factor-1 (HNF-1), a liver-enriched transcription factor implicated in the regulation of a number of genes in liver, kidney, stomach, intestine, and pancreas. Subsequent mobility shift assays confirmed that HNF-1alpha isoform was the predominant factor interacting with this region of the promoter. Mutation of the HNF-1 site resulted in a dramatic reduction in SRC promoter activity. Cotransfection studies demonstrated the promoter could be strongly transactivated by the HNF-1alpha isoform but not by the related HNF-1beta factor. Consistent with these results, we demonstrated that transcripts originating from the SRC1alpha promoter display a tissue restricted pattern of expression with highest levels present in stomach, kidney, and pancreas. These results indicate that SRC transcriptional regulation is much more complex than previously realized and implicates HNF-1 in both the tissue-specific regulation of the SRC gene in normal tissues and the overexpression of c-Src in certain human cancers.


Sujet(s)
Protéines de liaison à l'ADN , Régulation de l'expression des gènes/physiologie , Protéines nucléaires , Régions promotrices (génétique) , Protéines proto-oncogènes pp60(c-src)/génétique , Facteurs de transcription/physiologie , Séquence nucléotidique , ADN , Facteur nucléaire hépatocytaire HNF-1 , Facteur nucléaire hépatocytaire HNF-1 alpha , Facteur nucléaire hépatocytaire HNF-1 bêta , Humains , Données de séquences moléculaires , Liaison aux protéines , Proto-oncogène Mas , Facteurs de transcription/métabolisme , Transfection , Cellules cancéreuses en culture
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