Lung Tumor Microphysiological System with 3D Endothelium to Evaluate Modulators of T-Cell Infiltration

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract Lung cancer is a leading cause of death worldwide, with only a fraction of patients responding to immunotherapy. The correlation between increased T-cell infiltration and positive patient outcomes has motivated the search for therapeutics promoting T-cell infiltration. While transwell and spheroid platforms have been employed, these models lack flow and endothelial barriers, and cannot faithfully model T-cell adhesion, extravasation and migration through 3D tissue. Presented here is a 3D chemotaxis assay, in a lung tumor on chip model with 3D endothelium (LToC-Endo), to address this need. The described assay consists of a vascular tubule cultured under rocking flow, through which T-cells are added; a collagenous stromal barrier, through which T-cells infiltrate; and a chemoattractant/tumor compartment. Here, activated T-cells extravasate and infiltrate in response to gradients of rhCXCL11 and rhCXCL12. Adopting a T-cell activation protocol with a rest period enables proliferative burst prior to introducing T-cells into chips, increases T-cell expression of CXCR3 and CXCR4 receptors, and enhances assay sensitivity. In addition, incorporating this rest recovers endothelial activation in response to rhCXCL12. As a final control, we show that blocking ICAM-1 interferes with T-cell adhesion and chemotaxis. This microphysiological system, which mimics in vivo stromal and vascular barriers, can be used to evaluate potentiation of immune chemotaxis into tumors while probing for vascular responses to potential therapeutics. Finally, we propose a translational strategy by which this assay could be linked to preclinical and clinical models to support human dose prediction, personalized medicine, and the reduction, refinement, and replacement of animal models.

Authors 13

1. Katrina M. Wisdom corresponding

   GlaxoSmithKline (United States)

   Affiliation as printed

   In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA, USA

   In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 250 S. Collegeville Rd, Collegeville, PA, USA

2. Johnny Suijker Aachen Mimetas B. V

   Mimetas (Netherlands)

   Affiliation as printed

   Mimetas B.V., Biopartner Building 5, De Limes 7, 2342 DH Oegstgeest, The Netherlands

   Mimetas B.V., Biopartner Building 5, De Limes , 2342 DH Oegstgeest, The Netherlands

3. Lenie van den Broek Aachen Mimetas B. V

   Mimetas (Netherlands)

   Affiliation as printed

   Mimetas B.V., Biopartner Building 5, De Limes 7, 2342 DH Oegstgeest, The Netherlands

   Mimetas B.V., Biopartner Building 5, De Limes , 2342 DH Oegstgeest, The Netherlands

4. BanuPriya Sridharan

   GlaxoSmithKline (United States)

   Affiliation as printed

   In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA, USA

   In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 250 S. Collegeville Rd, Collegeville, PA, USA

5. Taraka Sai Pavan Grandhi

   GlaxoSmithKline (United States)

   Affiliation as printed

   In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA, USA

   In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 250 S. Collegeville Rd, Collegeville, PA, USA

6. Mahdi Lamb

   GlaxoSmithKline (United Kingdom)

   Affiliation as printed

   Genomic Sciences, Research, Pharmaceutical R&D, GSK, Gunnels Wood Rd, Stevenage SG1 2NY, UK

7. Steven A. Titus

   GlaxoSmithKline (United States)

   Affiliation as printed

   Genomic Sciences, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA USA

   Genomic Sciences, Research, Pharmaceutical R&D, GSK, 150 S. Collegeville Rd, Collegeville, PA USA

8. Andrew Gehman

   GlaxoSmithKline (United States)

   Affiliation as printed

   Development Biostatistics, Development, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA USA

9. Derek Poore

   GlaxoSmithKline (United States)

   Affiliation as printed

   Immuno-Oncology, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA USA

10. Niyant Shah

    GlaxoSmithKline (United States)

    Affiliation as printed

    Immuno-Oncology, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA USA

11. Shih‐Hsun Cheng

    GlaxoSmithKline (United States)

    Affiliation as printed

    In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA, USA

    In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 250 S. Collegeville Rd, Collegeville, PA, USA

12. Edward Jae-Hoon Kim

    GlaxoSmithKline (United States)

    Affiliation as printed

    Immuno-Oncology, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA USA

13. Jason E. Ekert

    GlaxoSmithKline (United States)

    Affiliation as printed

    In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 1250 S. Collegeville Rd, Collegeville, PA, USA

    In Vitro In Vivo Translation, Research, Pharmaceutical R&D, GSK, 250 S. Collegeville Rd, Collegeville, PA, USA