Rationale Longitudinal molecular characterisation may improve prognostic stratification in patients with biliary tract cancers (BTC), but the feasibility of serial sampling and the prognostic significance of genomic alterations detected before treatment remain incompletely defined. This interim analysis has two main objectives: to assess the feasibility of prospective sampling and molecular analysis in the early-stage and advanced-stage BTC cohorts and, on an exploratory basis, to evaluate the potential prognostic value of baseline whole-exome sequencing (WES) profiling in relation to clinical outcomes. Materials and methods The analysis included 20 patients who underwent radical surgery, most of whom were candidates for adjuvant capecitabine, and 22 patients with unresectable locally advanced or metastatic disease who received first-line cisplatin–gemcitabine combined with durvalumab or pembrolizumab. Baseline was defined as the time before the initiation of adjuvant therapy in the early-stage cohort and before chemoimmunotherapy in the advanced-stage cohort. The endpoints assessed were disease-free survival (DFS) and overall survival (OS) in the early-stage cohort, and progression-free survival (PFS) and OS in the advanced-stage cohort. Kaplan–Meier curves were compared using the log-rank test. The composite WES indicator combined pathogenic/likely pathogenic (P/LP) germline variants and/or the reported tumour tissue alterations. Longitudinal ctDNA/dPCR results were not available at the data cut-off of 31 July 2026. Results In the early-stage cohort, 93 blood sampling time points intended for the ctDNA programme were documented, with blood-based germline WES available in 16/20 patients and tumour tissue WES in 10/20 patients. In the advanced-stage cohort, 82 blood sampling time points were documented among the 26 patients evaluable for longitudinal assessment; within the primary analysis population of 22 patients, blood-based germline WES was available in 11/22 patients and tumour tissue WES in 2/22 patients. In the early-stage cohort, median DFS was 19.5 months (95% CI 13.9–NR), while median OS was not reached. In the advanced-stage cohort, median PFS and OS were 7.5 months (95% CI 4.8–9.9) and 12.1 months (95% CI 7.8–28.1), respectively. No statistically significant associations were observed between the composite baseline WES indicator and DFS or OS in the early-stage cohort, or between the same indicator and PFS or OS in the advanced-stage cohort. Nominally significant clinical associations, including those involving pT/pN and sex in the early-stage cohort and M stage/pulmonary metastases in the advanced-stage cohort, should be interpreted with caution because of the small sample size, multiple comparisons, or potential bias. Conclusions The prospective sample collection programme is operational, although molecular coverage and temporal completeness remain uneven. This analysis did not demonstrate a prognostic value for the composite baseline WES indicator; however, this finding does not rule out potential gene-specific effects. Molecular alterations and dynamics detected at subsequent time points will be assessed following completion of the longitudinal ctDNA/dPCR analyses and further maturation of follow-up. Keywords: biliary tract cancers; CCA; ctDNA; MRD; WES; chemoimmunotherapy.
Razionale La caratterizzazione molecolare longitudinale potrebbe migliorare la stratificazione prognostica nei pazienti affetti da neoplasie delle vie biliari, ma la fattibilità del campionamento seriato e il significato prognostico delle alterazioni genomiche rilevate prima del trattamento non sono del tutto definiti. La presente analisi ad interim ha due obiettivi principali: valutare la fattibilità del percorso di campionamento e di analisi molecolare prospettica nelle coorti BTC precoce e avanzata e, in via esplorativa, valutare il possibile valore prognostico della profilazione WES al basale rispetto agli outcome clinici. Materiali e metodi Sono stati analizzati 20 pazienti sottoposti a chirurgia radicale, candidati prevalentemente a terapia adiuvante con capecitabina, e 22 pazienti con malattia localmente avanzata non resecabile o metastatica, trattati in prima linea con cisplatino–gemcitabina associati a durvalumab o pembrolizumab. Il basale è stato definito prima dell’avvio della terapia adiuvante nella coorte precoce e prima della chemoimmunoterapia nella coorte avanzata. Gli endpoint considerati erano DFS e OS nella coorte precoce e PFS e OS nella coorte avanzata. Le curve di Kaplan–Meier sono state confrontate mediante test log-rank. L’indicatore WES composito aggregava varianti germinali P/LP e/o le alterazioni tissutali riportate. Gli esiti longitudinali di ctDNA/dPCR non erano disponibili al data cut-off del 31 luglio 2026. Risultati Nella coorte precoce sono stati documentati 93 timepoint ematici destinati al programma ctDNA, oltre al WES su sangue germinale in 16/20 pazienti e al WES su tessuto tumorale in 10/20. Nella coorte avanzata, nei 26 pazienti valutabili longitudinalmente sono stati documentati 82 timepoint ematici; nella popolazione analitica primaria di 22 pazienti, il WES su sangue germinale era disponibile in 11/22 pazienti e il WES su tessuto tumorale in 2/22 pazienti. Nella coorte precoce, la DFS mediana è stata di 19,5 mesi (IC95% 13,9–NR), mentre l’OS mediana non è stata raggiunta; nella coorte avanzata, la PFS e l’OS mediane sono risultate rispettivamente di 7,5 mesi (IC95% 4,8–9,9) e 12,1 mesi (IC95% 7,8–28,1). Non sono emerse associazioni statisticamente significative tra l’indicatore WES basale composito e la DFS o l’OS nella coorte precoce, né tra lo stesso indicatore e la PFS o l’OS nella coorte avanzata. Le associazioni cliniche nominalmente significative, incluse quelle relative a pT/pN e sesso nella coorte precoce e a stadio M/metastasi polmonari nella coorte avanzata, devono essere interpretate con cautela per la ridotta numerosità campionaria, la molteplicità dei confronti o la possibile presenza di bias. Conclusioni Il programma di raccolta prospettica risulta operativo, sebbene la copertura molecolare e la completezza temporale siano ancora disomogenee. L’analisi non ha documentato un valore prognostico dell’indicatore WES basale composito; tale risultato, tuttavia, non consente di escludere possibili effetti gene-specifici. Le alterazioni e le dinamiche molecolari rilevate ai timepoint successivi saranno valutate dopo il completamento delle analisi longitudinali del ctDNA/dPCR e un’ulteriore maturazione del follow-up. Keywords: neoplasie delle vie biliari; CCA; ctDNA; MRD; WES; chemoimmunoterapia.
Fattibilità del campionamento molecolare prospettico e valore prognostico della profilazione WES al basale in pazienti affetti da neoplasie delle vie biliari in stadio precoce e avanzato: analisi ad interim dello studio MRD-CCA/HCC.
MANASRA, ZEINAB
2025/2026
Abstract
Rationale Longitudinal molecular characterisation may improve prognostic stratification in patients with biliary tract cancers (BTC), but the feasibility of serial sampling and the prognostic significance of genomic alterations detected before treatment remain incompletely defined. This interim analysis has two main objectives: to assess the feasibility of prospective sampling and molecular analysis in the early-stage and advanced-stage BTC cohorts and, on an exploratory basis, to evaluate the potential prognostic value of baseline whole-exome sequencing (WES) profiling in relation to clinical outcomes. Materials and methods The analysis included 20 patients who underwent radical surgery, most of whom were candidates for adjuvant capecitabine, and 22 patients with unresectable locally advanced or metastatic disease who received first-line cisplatin–gemcitabine combined with durvalumab or pembrolizumab. Baseline was defined as the time before the initiation of adjuvant therapy in the early-stage cohort and before chemoimmunotherapy in the advanced-stage cohort. The endpoints assessed were disease-free survival (DFS) and overall survival (OS) in the early-stage cohort, and progression-free survival (PFS) and OS in the advanced-stage cohort. Kaplan–Meier curves were compared using the log-rank test. The composite WES indicator combined pathogenic/likely pathogenic (P/LP) germline variants and/or the reported tumour tissue alterations. Longitudinal ctDNA/dPCR results were not available at the data cut-off of 31 July 2026. Results In the early-stage cohort, 93 blood sampling time points intended for the ctDNA programme were documented, with blood-based germline WES available in 16/20 patients and tumour tissue WES in 10/20 patients. In the advanced-stage cohort, 82 blood sampling time points were documented among the 26 patients evaluable for longitudinal assessment; within the primary analysis population of 22 patients, blood-based germline WES was available in 11/22 patients and tumour tissue WES in 2/22 patients. In the early-stage cohort, median DFS was 19.5 months (95% CI 13.9–NR), while median OS was not reached. In the advanced-stage cohort, median PFS and OS were 7.5 months (95% CI 4.8–9.9) and 12.1 months (95% CI 7.8–28.1), respectively. No statistically significant associations were observed between the composite baseline WES indicator and DFS or OS in the early-stage cohort, or between the same indicator and PFS or OS in the advanced-stage cohort. Nominally significant clinical associations, including those involving pT/pN and sex in the early-stage cohort and M stage/pulmonary metastases in the advanced-stage cohort, should be interpreted with caution because of the small sample size, multiple comparisons, or potential bias. Conclusions The prospective sample collection programme is operational, although molecular coverage and temporal completeness remain uneven. This analysis did not demonstrate a prognostic value for the composite baseline WES indicator; however, this finding does not rule out potential gene-specific effects. Molecular alterations and dynamics detected at subsequent time points will be assessed following completion of the longitudinal ctDNA/dPCR analyses and further maturation of follow-up. Keywords: biliary tract cancers; CCA; ctDNA; MRD; WES; chemoimmunotherapy.| File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12075/28549