03340nas a2200841   4500000000100000000000100001008004100002260001500043653001200058653001800070653002500088653001100113653001100124653001800135653000900153653000900162653002400171653002600195653002200221653002800243653001400271653000900285653000800294653003100302653002000333653000800353653002700361653001100388653002500399653001800424653001300442653002400455653003400479100001800513700001500531700001500546700002200561700002700583700001400610700001600624700002000640700002400660700001900684700002100703700001700724700002100741700001300762700002000775700002100795700002200816700002300838700001800861700001900879700001300898700001800911700003000929700002800959700001900987700001901006700002001025700001901045700001901064700001901083700002001102700002201122700001801144700001801162245005901180300001801239490000801257520121901265022001402484       2018                            d  c2018-12-1310aAnimals10aB7-H1 Antigen10aCoculture Techniques10aFemale10aHumans10aImmunotherapy10aMale10aMice10aMice, Inbred BALB C10aModels, Immunological10aNeoplasm Proteins10aNeoplasms, Experimental10aorganoids10aPD-110aPDO10aReceptors, Antigen, T-Cell10aT cell receptor10aTCR10aTumor Microenvironment10aCancer10acheckpoint inhibitor10aImmunotherapy10aorganoid10asingle-cell RNA-seq10atumor-infiltrating lymphocyte1 aJames T. Neal1 aXingnan Li1 aJunjie Zhu1 aValeria Giangarra1 aCaitlin L. Grzeskowiak1 aJihang Ju1 aIris H. Liu1 aShin-Heng Chiou1 aAmeen A. Salahudeen1 aAmber R. Smith1 aBrian C. Deutsch1 aLillian Liao1 aAllison J. Zemek1 aFan Zhao1 aKasper Karlsson1 aLiora M. Schultz1 aThomas J. Metzner1 aLincoln D. Nadauld1 aYuen-Yi Tseng1 aSahar Alkhairy1 aCoyin Oh1 aPaula Keskula1 aDaniel Mendoza-Villanueva1 aFrancisco M. De La Vega1 aPamela L. Kunz1 aJoseph C. Liao1 aJohn T. Leppert1 aJohn B. Sunwoo1 aChiara Sabatti1 aJesse S. Boehm1 aWilliam C. Hahn1 aGrace X. Y. Zheng1 aMark M. Davis1 aCalvin J. Kuo00aOrganoid Modeling of the Tumor Immune Microenvironment  a1972-1988.e160 v1753 aIn vitro cancer cultures, including three-dimensional organoids, typically contain exclusively neoplastic epithelium but require artificial reconstitution to recapitulate the tumor microenvironment (TME). The co-culture of primary tumor epithelia with endogenous, syngeneic tumor-infiltrating lymphocytes (TILs) as a cohesive unit has been particularly elusive. Here, an air-liquid interface (ALI) method propagated patient-derived organoids (PDOs) from >100 human biopsies or mouse tumors in syngeneic immunocompetent hosts as tumor epithelia with native embedded immune cells (T, B, NK, macrophages). Robust droplet-based, single-cell simultaneous determination of gene expression and immune repertoire indicated that PDO TILs accurately preserved the original tumor T cell receptor (TCR) spectrum. Crucially, human and murine PDOs successfully modeled immune checkpoint blockade (ICB) with anti-PD-1- and/or anti-PD-L1 expanding and activating tumor antigen-specific TILs and eliciting tumor cytotoxicity. Organoid-based propagation of primary tumor epithelium en bloc with endogenous immune stroma should enable immuno-oncology investigations within the TME and facilitate personalized immunotherapy testing.  a1097-4172