Targeting Ion Channel Dysregulation in Colorectal Cancer: Identification of Clofilium as a Potent Inhibitor of Tumor Growth and Metastatic Potential
Abstract
Colorectal cancer (CRC) remains a major cause of cancer mortality, motivating strategies targeting noncanonical tumor vulnerabilities. Public gene-expression datasets identified ion channel–associated genes upregulated in primary CRC and COLO 205 cells versus normal colon, guiding a screen of 17 ion-modulating compounds. Nine inhibited COLO 205 proliferation. Leading compounds suppressed COLO 205, HT-29, and Caco-2 growth, predominantly inducing cytostasis with G1/G0 accumulation. Most were less active in 3D COLO 205 spheroids than 2D monolayers; only compounds blocking both potassium and calcium channels reduced spheroid size. However, our lead compound, clofilium retained 3D activity, reducing COLO 205 and HT-29 spheroid size, cohesion, migration, and invasion, plus COLO 205 intravasation and endothelial sprouting in heterotypic tumor–stroma spheroids. Across leading compounds, cytostasis frequently coincided with prolonged depolarization and reduced cytoplasmic Ca2+. Clofilium induced sustained depolarization, growth suppression after withdrawal, senescence-associated features, and altered mitochondrial membrane potential and content. RNA sequencing of clofilium-treated COLO 205 cells revealed suppressed DNA-replication, cell-cycle, chromatin-maintenance, CRC-stemness programs, and induced ER-stress, cell-cycle-inhibitory, metabolic-adaptation, innate immune, and ion-homeostasis pathways. Protein analyses in COLO 205 and HT-29 cells showed increased NDRG1, p21, IRE1α, and CHOP, with cell-line-specific kinase-signaling changes. In COLO 205 xenografts, daily intravenous clofilium reduced mean tumor-volume change from baseline by 43% versus vehicle controls, with limited observable systemic toxicity. These findings implicate CRC ion-transport networks and bioelectric states as therapeutic vulnerabilities. Clofilium emerged as a lead ionoceutical candidate with activity across 2D, 3D, heterotypic microenvironment, and xenograft models, supporting further evaluation of multi-channel ion-modulating strategies for CRC.
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BibTeX
@misc{mathews2026targeting,
title = {{Targeting Ion Channel Dysregulation in Colorectal Cancer: Identification of Clofilium as a Potent Inhibitor of Tumor Growth and Metastatic Potential}},
author = {Juanita Mathews and Sydney A. Palmer and Sammy Hansali and Jordan Strasser and Alyssa Liao and Gayathri Kondakath and Zoe Mozden Weiner and Léo Pio-Lopez and Franz Kuchling and Eva Swei},
howpublished = {OSF Preprints},
year = {2026},
month = oct,
doi = {10.31219/osf.io/w5dq7_v4},
url = {https://osf.io/w5dq7_v4/},
note = {CC BY 4.0},
}
RIS
TY - UNPB
TI - Targeting Ion Channel Dysregulation in Colorectal Cancer: Identification of Clofilium as a Potent Inhibitor of Tumor Growth and Metastatic Potential
AU - Juanita Mathews
AU - Sydney A. Palmer
AU - Sammy Hansali
AU - Jordan Strasser
AU - Alyssa Liao
AU - Gayathri Kondakath
AU - Zoe Mozden Weiner
AU - Léo Pio-Lopez
AU - Franz Kuchling
AU - Eva Swei
PY - 2026
DA - 2026/10/19
PB - OSF Preprints
DO - 10.31219/osf.io/w5dq7_v4
UR - https://osf.io/w5dq7_v4/
AB - Colorectal cancer (CRC) remains a major cause of cancer mortality, motivating strategies targeting noncanonical tumor vulnerabilities. Public gene-expression datasets identified ion channel–associated genes upregulated in primary CRC and COLO 205 cells versus normal colon, guiding a screen of 17 ion-modulating compounds. Nine inhibited COLO 205 proliferation. Leading compounds suppressed COLO 205, HT-29, and Caco-2 growth, predominantly inducing cytostasis with G1/G0 accumulation. Most were less active in 3D COLO 205 spheroids than 2D monolayers; only compounds blocking both potassium and calcium channels reduced spheroid size. However, our lead compound, clofilium retained 3D activity, reducing COLO 205 and HT-29 spheroid size, cohesion, migration, and invasion, plus COLO 205 intravasation and endothelial sprouting in heterotypic tumor–stroma spheroids. Across leading compounds, cytostasis frequently coincided with prolonged depolarization and reduced cytoplasmic Ca2+. Clofilium induced sustained depolarization, growth suppression after withdrawal, senescence-associated features, and altered mitochondrial membrane potential and content. RNA sequencing of clofilium-treated COLO 205 cells revealed suppressed DNA-replication, cell-cycle, chromatin-maintenance, CRC-stemness programs, and induced ER-stress, cell-cycle-inhibitory, metabolic-adaptation, innate immune, and ion-homeostasis pathways. Protein analyses in COLO 205 and HT-29 cells showed increased NDRG1, p21, IRE1α, and CHOP, with cell-line-specific kinase-signaling changes. In COLO 205 xenografts, daily intravenous clofilium reduced mean tumor-volume change from baseline by 43% versus vehicle controls, with limited observable systemic toxicity. These findings implicate CRC ion-transport networks and bioelectric states as therapeutic vulnerabilities. Clofilium emerged as a lead ionoceutical candidate with activity across 2D, 3D, heterotypic microenvironment, and xenograft models, supporting further evaluation of multi-channel ion-modulating strategies for CRC.
ER -