This U01 is in response to FOA PAR-16-105 Cancer Tissue Engineering Collaborative: Enabling Biomimetic Tissue-Engineered Technologies for Cancer Research. In it we will address the substantial gap in our knowledge of the initiation and progression of metastatic colorectal cancer (CRC). This gap is due, in large part, to the fact that current in vitro, ex vivo, and even in vivo research models are hard to establish or have limited applicability. Accordingly, our objectives are to show the utility of three innovative models of metastasis (a long-term organotypic model and a model that uses multi-organ microfluidic devices) and a new in vivo model (immunocompetent blastocyst model). Using these models, we will investigate the interactive cellular secretome, differentiation, migration and invasion of the primary and metastatic fibroblast tumor microenvironment in order to identify critical contributions to CRC metastasis. Our hypothesis is: inflammation-associated fibroblasts and altered metabolic conditions promote epithelial metastasis via transcriptional regulation. Based on our recent successful engineering of novel models, we propose 3 Aims.
Aim 1. To integrate cytokine-secreting stroma into our organotypic model in order to study the influence of inflammation on cancer stem cells (SCs) and on invasion/migration cell phenotypes.
Aim 2. To engineer a liver metastasis model using a microfluidic body-on-a- chip platform to study transcriptome and epigenetic reprogramming of metastatic cells.
Aim 3. To validate our in vitro models using in vivo immunoproficient murine metastatic models.
In Aim 1, decellularized human colons will be repopulated with normal or cancerous fibroblasts from the stroma, and cancer epithelial cells in the form of cancer SCs and organoids to ask how these manipulations influence invasion and differentiation in this model. To complement the cellular findings, we will identify alterations in the transcriptome using RNA-seq and ATAC- seq.
Aim 2 uses the innovative body-on-a chip microfluidic device to test autocrine and paracrine proliferative and secretory responses under conditions that favor oncogenic progression, including hypoxia and low glucose. As in Aim 1, epithelia and fibroblasts in the different milieus will be transcriptionally profiled.
Aim 3 uses a new immunocompetent blastocyst model and will undergo the same transcriptional profiling. All 3 models will be linked using comparative, informatics approaches. The approach is innovative as it uses human colon tissues in all Aims and asks about the transcriptional contribution and landscape of accessible chromatin in each model. Our multidisciplinary team includes a colorectal surgeon-scientist and biomedical engineers with expertise in cancer SCs, signaling, metastases, and organotypic and organ-on-a-chip platforms. The findings will be significant, as they will establish improved approaches for studying the pathogenesis of advanced CRC, and for developing patient-derived, high-throughput cell models to test therapeutic interventions.

Public Health Relevance

The proposed research is relevant to public health because understanding the pathogenesis of colorectal cancer metastases will result in the development of innovative preventive and therapeutic strategies. The research is relevant to the part of the NIH's mission that addresses expansion of knowledge to enhance health and to reduce illness in this context. By preventing or treating colorectal cancer metastases, this proposal will also address that part of the NIH's mission to lengthen life.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01CA214300-03
Application #
9766204
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Zahir, Nastaran Z
Project Start
2017-09-25
Project End
2022-08-30
Budget Start
2019-09-01
Budget End
2020-08-31
Support Year
3
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Cleveland Clinic Lerner
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
135781701
City
Cleveland
State
OH
Country
United States
Zip Code
44195
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Signs, Steven A; Fisher, Robert C; Tran, Uyen et al. (2018) Stromal miR-20a controls paracrine CXCL8 secretion in colitis and colon cancer. Oncotarget 9:13048-13059
Chen, Kai-Yuan; Srinivasan, Tara; Lin, Christopher et al. (2018) Single-Cell Transcriptomics Reveals Heterogeneity and Drug Response of Human Colorectal Cancer Organoids. Conf Proc IEEE Eng Med Biol Soc 2018:2378-2381
Wang, Ying I; Shuler, Michael L (2018) UniChip enables long-term recirculating unidirectional perfusion with gravity-driven flow for microphysiological systems. Lab Chip 18:2563-2574
Shawki, Sherief; Ashburn, Jean; Signs, Steven A et al. (2018) Colon Cancer: Inflammation-Associated Cancer. Surg Oncol Clin N Am 27:269-287