Kenneth K. Lee

Overview of PDO and TIL analyses

Gating strategy for the phenotyping of TIL cultures at REP day 14 by spectral cytometry

Fluorochrome-conjugated antibodies

Patient demographic and clinicopathological characteristics

Purpose:Unsatisfactory outcomes in pancreatic ductal adenocarcinoma (PDAC) highlight the need to identify precision-based treatment modalities. We aimed to utilize patient-derived organoids (PDO) to predict the most effective individual components of potential combination therapies.Experimental Design:Autologous αβ- and γδ-enriched tumor-infiltrating lymphocyte (TIL) and PDO cultures were established from resected PDAC tissue. PDOs were characterized by immunostaining, targeted next-generation sequencing, and drug screening with standard-of-care multidrug chemotherapeutic regimens. Expanded TIL cultures were profiled through flow cytometry, sequencing, and potency against autologous PDOs.Results:PDO cultures were established with an 80% success rate. In addition to faithfully recapitulating molecular and histologic features of the parental tumor, PDOs were sufficiently expanded for pharmacotyping within a clinically relevant time frame of 36 days. Notably, PDO chemotherapeutic sensitivity profiles correlated with patient serum CA 19-9 dynamics and recurrence-free survival. Furthermore, expanded γδ-enriched TILs supported the activation of αβ TCR+ cells and demonstrated more potent functionality in response to autologous PDO targets. Importantly, infiltration of γδ T cells within pancreatic tumor tissue was associated with improved overall survival.Conclusions:We confirm the feasibility of generating PDOs within a clinically relevant time frame and provide evidence of their utility for advancing therapeutic success in PDAC.

Schematic illustrating the simultaneous expansion of autologous TIL and PDOs from PDAC tissue

Therapeutic profiling of PDOs unveils variability in drug responses and interactions

γδ-enriched TIL directly and indirectly contribute to functional antitumor immune responses

PDOs from biopsy specimens demonstrate greater chemotherapeutic sensitivity

Purpose: Unsatisfactory outcomes in pancreatic ductal adenocarcinoma (PDAC) highlight the need to identify precision-based treatment modalities. We aimed to utilize patient-derived organoids (PDO) to predict the most effective individual components of potential combination therapies. Experimental Design: Autologous αβ- and γδ-enriched tumor-infiltrating lymphocyte (TIL) and PDO cultures were established from resected PDAC tissue. PDOs were characterized by immunostaining, targeted next-generation sequencing, and drug screening with standard-of-care multidrug chemotherapeutic regimens. Expanded TIL cultures were profiled through flow cytometry, sequencing, and potency against autologous PDOs. Results: PDO cultures were established with an 80% success rate. In addition to faithfully recapitulating molecular and histologic features of the parental tumor, PDOs were sufficiently expanded for pharmacotyping within a clinically relevant time frame of 36 days. Notably, PDO chemotherapeutic sensitivity profiles correlated with patient serum CA 19-9 dynamics and recurrence-free survival. Furthermore, expanded γδ-enriched TILs supported the activation of αβ TCR+ cells and demonstrated more potent functionality in response to autologous PDO targets. Importantly, infiltration of γδ T cells within pancreatic tumor tissue was associated with improved overall survival. Conclusions: We confirm the feasibility of generating PDOs within a clinically relevant time frame and provide evidence of their utility for advancing therapeutic success in PDAC.