Peristalsis-Associated Mechanotransduction Drives Malignant Progression of Colorectal Cancer

Clevenger, Abigail J.; McFarlin, Maygan K.; Collier, Claudia A.; Sheshadri, Vibha S.; Madyastha, Anirudh K.; Gorley, John Paul M.; Solberg, Spencer C.; Stratman, Amber N.; Raghavan, Shreya A.

doi:10.1007/s12195-023-00776-w

Cited by 4 publications

(13 citation statements)

References 114 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Polydimethylsiloxane (PDMS) membranes were prepared at a 10:1 ratio using previously established protocols 25,26 . To maximize seeding of cells on PDMS, the cell seeding area of the membranes were coated with 200 µg/mL of Collagen I.…”

Section: Preparation Of Cell Seeded Membranes and Bioreactor Assemblymentioning

confidence: 99%

“…Operation of the peristalsis bioreactor was followed as previously reported 25,26 . Briefly, media was removed from each cell-seeded PDMS membrane.…”

Section: Preparation Of Cell Seeded Membranes and Bioreactor Assemblymentioning

confidence: 99%

“…The bioreactor was assembled using commercially available zip ties and connected to the peristalsis pump. The assembled set up was placed into the 37°C incubator and connected to an Arduino that ran a pre-programmed code with the following parameters: 0.4 Pa shear and 15% cyclic strain at 12 rpm 25,26 . Cells were incubated, either as static controls or stimulated in the peristalsis bioreactor at 37°C for 24 hours and then collected for downstream analysis.…”

Section: Preparation Of Cell Seeded Membranes and Bioreactor Assemblymentioning

confidence: 99%

“…In previous studies, we developed and validated a peristalsis bioreactor that mimics the forces associated with colonic peristalsis 25 . Using the peristalsis bioreactor, we demonstrated that HCT116 KRAS G13D cells exposed to peristalsis increased cancer stem cell markers relative to static culture 26 . HCT116 cells also developed invasive signatures downstream of peristalsis, indicating malignant progression.…”

Section: Introductionmentioning

confidence: 97%

“…Leveraging the peristalsis bioreactor, multiple cell lines and patient-derived xenograft lines were employed to test the hypothesis that activating KRAS mutations change mechanotransduction via cancer stem cell enrichment. Our focus was on the Leucine rich repeat containing G protein couple receptor, LGR5, a cancer stem cell marker which our previous work strongly demonstrated was downstream of Wnt activation 26 . Here, we characterize changes in the Wnt pathway as a function of KRAS mutation status in response to peristalsis.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

OncogenicKRASMutations Confer a Unique Mechanotransduction Response to Peristalsis in Colorectal Cancer Cells

Clevenger,

Collier,

Gorley

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

Colorectal cancer (CRC) tumors start as precancerous polyps on the inner lining of the colon or rectum, where they are exposed to the mechanics of colonic peristalsis. Our previous work leveraged a custom-built peristalsis bioreactor to demonstrate that colonic peristalsis led to cancer stem cell enrichment in colorectal cancer cells. However, this malignant mechanotransductive response was confined to select CRC lines that harbored an oncogenic mutation in the KRAS gene. In this work, therefore, we explored the involvement of activating KRAS mutations on peristalsis-associated mechanotransduction in CRC. Peristalsis enriched the cancer stem cell marker LGR5 in KRAS mutant (G13D, etc.) lines, in a Wnt-independent manner. Conversely, LGR5 enrichment in wild type KRAS lines exposed to peristalsis were minimal. LGR5 enrichment downstream of peristalsis translated to increased tumorigenicity in vivo in KRAS mutant vs. wild type lines. Differences in mechanotransduction response was additionally apparent via unbiased gene set enrichment analysis, where many unique pathways were enriched in wild type vs. mutant lines, in response to peristalsis. Interestingly, peristalsis also triggered β-catenin nuclear localization independent of Wnt, particularly in KRAS mutant lines. The central involvement of KRAS in the mechanotransductive responses was validated via gain and loss of function strategies. β-catenin activation and LGR5 enrichment downstream of peristalsis converged to the activation of the MEK/ERK kinase cascade, that remains active in cells that harbor oncogenic KRAS mutations. Taken together, our results demonstrated that oncogenic KRAS mutations conferred a unique peristalsis-associated mechanotransduction response to colorectal cancer cells, resulting in cancer stem cell enrichment and increased tumorigenicity. These mechanosensory connections can be leveraged in improving the sensitivity of emerging therapies that target oncogenic KRAS.

show abstract

Section: Preparation Of Cell Seeded Membranes and Bioreactor Assemblymentioning

confidence: 99%

“…Operation of the peristalsis bioreactor was followed as previously reported 25,26 . Briefly, media was removed from each cell-seeded PDMS membrane.…”

Section: Preparation Of Cell Seeded Membranes and Bioreactor Assemblymentioning

confidence: 99%

Section: Preparation Of Cell Seeded Membranes and Bioreactor Assemblymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

OncogenicKRASMutations Confer a Unique Mechanotransduction Response to Peristalsis in Colorectal Cancer Cells

Clevenger,

Collier,

Gorley

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Bridging barriers: advances and challenges in modeling biological barriers and measuring barrier integrity in organ-on-chip systems

Ugodnikov,

Persson,

Simmons

2024

Lab Chip

View full text Add to dashboard Cite

show abstract

Advances in cancer mechanobiology: Metastasis, mechanics, and materials

Clevenger,

McFarlin,

Gorley

et al. 2024

APL Bioengineering

Self Cite

View full text Add to dashboard Cite

Within the tumor microenvironment (TME), tumor cells are exposed to numerous mechanical forces, both internally and externally, which contribute to the metastatic cascade. From the initial growth of the tumor to traveling through the vasculature and to the eventual colonization of distant organs, tumor cells are continuously interacting with their surroundings through physical contact and mechanical force application. The mechanical forces found in the TME can be simplified into three main categories: (i) shear stress, (ii) tension and strain, and (iii) solid stress and compression. Each force type can independently impact tumor growth and progression. Here, we review recent bioengineering strategies, which have been employed to establish the connection between mechanical forces and tumor progression. While many cancers are explored in this review, we place great emphasis on cancers that are understudied in their response to mechanical forces, such as ovarian and colorectal cancers. We discuss the major steps of metastatic transformation and present novel, recent advances in model systems used to study how mechanical forces impact the study of the metastatic cascade. We end by summarizing systems that incorporate multiple forces to expand the complexity of our understanding of how tumor cells sense and respond to mechanical forces in their environment. Future studies would also benefit from the inclusion of time or the aspect of mechanical memory to further enhance this field. While the knowledge of mechanical forces and tumor metastasis grows, developing novel materials and in vitro systems are essential to providing new insight into predicting, treating, and preventing cancer progression and metastasis.

show abstract

Peristalsis-Associated Mechanotransduction Drives Malignant Progression of Colorectal Cancer

Cited by 4 publications

References 114 publications

OncogenicKRASMutations Confer a Unique Mechanotransduction Response to Peristalsis in Colorectal Cancer Cells

OncogenicKRASMutations Confer a Unique Mechanotransduction Response to Peristalsis in Colorectal Cancer Cells

Bridging barriers: advances and challenges in modeling biological barriers and measuring barrier integrity in organ-on-chip systems

Advances in cancer mechanobiology: Metastasis, mechanics, and materials

Contact Info

Product

Resources

About