ROR1-CAR T cells are effective against lung and breast cancer in advanced microphysiologic 3D tumor models

JCI Insight. 2019 Sep 19;4(18):e126345. doi: 10.1172/jci.insight.126345.


Solid tumors impose immunologic and physical barriers to the efficacy of chimeric antigen receptor (CAR) T cell therapy that are not reflected in conventional preclinical testing against singularized tumor cells in 2-dimensional culture. Here, we established microphysiologic three-dimensional (3D) lung and breast cancer models that resemble architectural and phenotypical features of primary tumors and evaluated the antitumor function of receptor tyrosine kinase-like orphan receptor 1-specific (ROR1-specific) CAR T cells. 3D tumors were established from A549 (non-small cell lung cancer) and MDA-MB-231 (triple-negative breast cancer) cell lines on a biological scaffold with intact basement membrane (BM) under static and dynamic culture conditions, which resulted in progressively increasing cell mass and invasive growth phenotype (dynamic > static; MDA-MB-231 > A549). Treatment with ROR1-CAR T cells conferred potent antitumor effects. In dynamic culture, CAR T cells actively entered arterial medium flow and adhered to and infiltrated the tumor mass. ROR1-CAR T cells penetrated deep into tumor tissue and eliminated multiple layers of tumor cells located above and below the BM. The microphysiologic 3D tumor models developed in this study are standardized, scalable test systems that can be used either in conjunction with or in lieu of animal testing to interrogate the antitumor function of CAR T cells and to obtain proof of concept for their safety and efficacy before clinical application.

Keywords: Breast cancer; Immunology; Immunotherapy; Lung cancer; Oncology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Testing Alternatives
  • Cell Culture Techniques / methods*
  • Cell Line, Tumor
  • Female
  • Humans
  • Immunotherapy, Adoptive / methods*
  • Lung Neoplasms / immunology
  • Lung Neoplasms / therapy*
  • Receptor Tyrosine Kinase-like Orphan Receptors / immunology*
  • Receptors, Chimeric Antigen / immunology*
  • Single-Chain Antibodies / immunology
  • Spheroids, Cellular
  • T-Lymphocytes / immunology
  • T-Lymphocytes / transplantation
  • Triple Negative Breast Neoplasms / immunology
  • Triple Negative Breast Neoplasms / therapy*


  • Receptors, Chimeric Antigen
  • Single-Chain Antibodies
  • ROR1 protein, human
  • Receptor Tyrosine Kinase-like Orphan Receptors