2. Academic Validation
  3. Tumor immune microenvironment reconstitution in patient-derived organoids enables therapy modeling for NSCLC

Tumor immune microenvironment reconstitution in patient-derived organoids enables therapy modeling for NSCLC

  • Cell Rep Methods. 2026 Jun 15;6(6):101339. doi: 10.1016/j.crmeth.2026.101339.
Enrique Podaza 1,  Jared Capuano 2,  Hui-Hsuan Kuo 2,  Majd Al Assaad 3,  Geoffrey Markowitz 4,  M Victoria Revuelta 5,  John Nguyen 2,  Adriana Irizarry 2,  Hiranmayi Ravichandran 2,  Sarah Ackermann 2,  Troy Kane 2,  Jyothi Manohar 2,  Alyssa Duren-Lubanski 2,  Michael Sigouros 2,  Jenna Moyer 2,  Bhavneet Bhinder 6,  Pooja Chandra 6,  Murtaza Malbari 4,  Karsten Boehnke 7,  Juan Miguel Mosquera 3,  Vivek Mittal 4,  Andrea Sboner 8,  Hamza Gokozan 9,  Nasser Altorki 4,  Olivier Elemento 10,  M Laura Martin 11
Affiliations

Affiliations

  • 1 Caryl and Israel Englander Institute for Precision Medicine, Weill Cornell Medicine, New York, NY 10021, USA; Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10021, USA.
  • 2 Caryl and Israel Englander Institute for Precision Medicine, Weill Cornell Medicine, New York, NY 10021, USA.
  • 3 Caryl and Israel Englander Institute for Precision Medicine, Weill Cornell Medicine, New York, NY 10021, USA; Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY 10021, USA.
  • 4 Cardiothoracic Surgery, Weill Cornell Medical College, New York, NY 10021, USA.
  • 5 Department of Hematology and Oncology, Weill Cornell Medical College, New York, NY 10021, USA.
  • 6 Caryl and Israel Englander Institute for Precision Medicine, Weill Cornell Medicine, New York, NY 10021, USA; Institute for Computational Biomedicine, Weill Cornell Medicine, New York, NY 10021, USA.
  • 7 Eli Lilly and Company, Lilly Oncology, Discovery Technologies, New York, NY 10016, USA.
  • 8 Caryl and Israel Englander Institute for Precision Medicine, Weill Cornell Medicine, New York, NY 10021, USA; Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY 10021, USA; Institute for Computational Biomedicine, Weill Cornell Medicine, New York, NY 10021, USA.
  • 9 Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY 10021, USA.
  • 10 Caryl and Israel Englander Institute for Precision Medicine, Weill Cornell Medicine, New York, NY 10021, USA; Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY 10021, USA; Institute for Computational Biomedicine, Weill Cornell Medicine, New York, NY 10021, USA.
  • 11 Caryl and Israel Englander Institute for Precision Medicine, Weill Cornell Medicine, New York, NY 10021, USA. Electronic address: [email protected].
PMID: 42134319 DOI: 10.1016/j.crmeth.2026.101339
Abstract

Non-small cell lung Cancer (NSCLC) remains a leading cause of cancer-related mortality. Despite various therapeutic options, treatment resistance is common, underscoring the need for effective combination therapies and reliable pre-clinical models for patient-specific evaluation. Here, we describe strategies for reconstituting tumor immune microenvironment (TIME) components within patient-derived tumor Organoid (PDTO) cultures. We established a tumor processing pipeline that enables concurrent expansion of tumor-infiltrating lymphocytes (TILs) and PDTO generation from the same resection. We optimized scalable assays to assess IFN-γ secretion and T cell cytotoxicity with immune checkpoint inhibitors (alone or in combination) and targeted inhibitors, capturing inter-patient heterogeneity and intra-patient variations between TILs and peripheral blood mononuclear cells (PBMCs). Additionally, we developed methods for differentiating PDTO-specific tumor-associated macrophages (TAMs) and established PDTO-TAM co-culture systems to evaluate TAM effects on PDTO growth and chemotherapy sensitivity. All approaches are scalable to high-throughput levels, highlighting the value of TIME-PDTO co-cultures for therapeutic modeling and precision medicine.

Keywords

CP: cancer biology; CP: immunology; T cells; co-cultures; immunotherapy; tumor microenviroment; tumor organoids; tumor-infiltrating lymphocytes.

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