American Association for Cancer Research
Browse
00085472can191523-sup-222308_3_supp_6255665_q9b184.pdf (13.65 MB)

Supplementary Figure 4 from Pattern of Invasion in Human Pancreatic Cancer Organoids Is Associated with Loss of SMAD4 and Clinical Outcome

Download (13.65 MB)
journal contribution
posted on 2023-03-31, 03:21 authored by Wenjie Huang, Bernat Navarro-Serer, Yea Ji Jeong, Peter Chianchiano, Limin Xia, Claudio Luchini, Nicola Veronese, Cameron Dowiak, Tammy Ng, Maria A. Trujillo, Bo Huang, Michael J. Pflüger, Anne M. Macgregor-Das, Gemma Lionheart, Danielle Jones, Kohei Fujikura, Kim-Vy Nguyen-Ngoc, Neil M. Neumann, Vincent P. Groot, Alina Hasanain, A. Floortje van Oosten, Sandra E. Fischer, Steven Gallinger, Aatur D. Singhi, Amer H. Zureikat, Randall E. Brand, Matthias M. Gaida, Stefan Heinrich, Richard A. Burkhart, Jin He, Christopher L. Wolfgang, Michael G. Goggins, Elizabeth D. Thompson, Nicholas J. Roberts, Andrew J. Ewald, Laura D. Wood

Impact of Culture Matrix on PDAC Organoid Invasion

Funding

American Cancer Society

NIH

NCI

NIDDK

Sol Goldman Pancreatic Cancer Research Center

AGA-Bernard Lee Schwartz Foundation Research

Pancreatic Cancer

Sidney Kimmel Foundation for Cancer Research

AACR-Incyte Corporation Career Development

Pancreatic Cancer Research

Joseph C Monastra Foundation

Gerald O Mann Charitable Foundation

Ontario Institute for Cancer Research

Government of Ontario

Princess Margaret Cancer Foundation

Terry Fox Research Institute

Canadian Cancer Society Research Institute

History

ARTICLE ABSTRACT

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy characterized by extensive local invasion and systemic spread. In this study, we employed a three-dimensional organoid model of human pancreatic cancer to characterize the molecular alterations critical for invasion. Time-lapse microscopy was used to observe invasion in organoids from 25 surgically resected human PDAC samples in collagen I. Subsequent lentiviral modification and small-molecule inhibitors were used to investigate the molecular programs underlying invasion in PDAC organoids. When cultured in collagen I, PDAC organoids exhibited two distinct, morphologically defined invasive phenotypes, mesenchymal and collective. Each individual PDAC gave rise to organoids with a predominant phenotype, and PDAC that generated organoids with predominantly mesenchymal invasion showed a worse prognosis. Collective invasion predominated in organoids from cancers with somatic mutations in the driver gene SMAD4 (or its signaling partner TGFBR2). Reexpression of SMAD4 abrogated the collective invasion phenotype in SMAD4-mutant PDAC organoids, indicating that SMAD4 loss is required for collective invasion in PDAC organoids. Surprisingly, invasion in passaged SMAD4-mutant PDAC organoids required exogenous TGFβ, suggesting that invasion in SMAD4-mutant organoids is mediated through noncanonical TGFβ signaling. The Rho-like GTPases RAC1 and CDC42 acted as potential mediators of TGFβ-stimulated invasion in SMAD4-mutant PDAC organoids, as inhibition of these GTPases suppressed collective invasion in our model. These data suggest that PDAC utilizes different invasion programs depending on SMAD4 status, with collective invasion uniquely present in PDAC with SMAD4 loss. Organoid models of PDAC highlight the importance of SMAD4 loss in invasion, demonstrating that invasion programs in SMAD4-mutant and SMAD4 wild-type tumors are different in both morphology and molecular mechanism.

Usage metrics

    Cancer Research

    Licence

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC