Microtentacle Formation in Ovarian Carcinoma
- Creators
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Reader, Jocelyn C.
- Fan, Cong
- Ory, Eleanor Claire-Higgins
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Ju, Julia
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Lee, Rachel
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Vitolo, Michele I.
- Smith, Paige
- Wu, Sulan
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Ching, Mc Millan Nicol
- Asiedu, Emmanuel B.
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Jewell, Christopher M.
- Rao, Gautam G.
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Fulton, Amy
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Webb, Tonya J.
- Yang, Peixin
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Santin, Alessandro D.
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Huang, Huang-Chiao
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Martin, Stuart S.
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Roque, Dana M.
Abstract
Background: The development of chemoresistance to paclitaxel and carboplatin represents a major therapeutic challenge in ovarian cancer, a disease frequently characterized by malignant ascites and extrapelvic metastasis. Microtentacles (McTNs) are tubulin-based projections observed in detached breast cancer cells. In this study, we investigated whether ovarian cancers exhibit McTNs and characterized McTN biology. Methods: We used an established lipid-tethering mechanism to suspend and image individual cancer cells. We queried a panel of immortalized serous (OSC) and clear cell (OCCC) cell lines as well as freshly procured ascites and human ovarian surface epithelium (HOSE). We assessed by Western blot β-tubulin isotype, α-tubulin post-translational modifications and actin regulatory proteins in attached/detached states. We studied clustering in suspended conditions. Effects of treatment with microtubule depolymerizing and stabilizing drugs were described. Results: Among cell lines, up to 30% of cells expressed McTNs. Four McTN morphologies (absent, symmetric-short, symmetric-long, tufted) were observed in immortalized cultures as well as ascites. McTN number/length varied with histology according to metastatic potential. Most OCCC overexpressed class III ß-tubulin. OCCC/OSC cell lines exhibited a trend towards more microtubule-stabilizing post-translational modifications of α-tubulin relative to HOSE. Microtubule depolymerizing drugs decreased the number/length of McTNs, confirming that McTNs are composed of tubulin. Cells that failed to form McTNs demonstrated differential expression of α-tubulin- and actin-regulating proteins relative to cells that form McTNs. Cluster formation is more susceptible to microtubule targeting agents in cells that form McTNs, suggesting a role for McTNs in aggregation. Conclusions: McTNs likely participate in key aspects of ovarian cancer metastasis. McTNs represent a new therapeutic target for this disease that could refine therapies, including intraperitoneal drug delivery.
Additional Information
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Received: 25 October 2021 / Revised: 16 January 2022 / Accepted: 19 January 2022 / Published: 4 February 2022. (This article belongs to the Special Issue Microtubule Dynamics and Cancer). This work was supported by the Department of Obstetrics, Gynecology & Reproductive Sciences at University of Maryland School of Medicine. JCR would like to acknowledge University of Maryland Eastern Shore for salary support during the completion of this manuscript. Author Contributions. Conceptualization: D.M.R., S.S.M.; Funding Acquisition: D.M.R.; Methodology: S.S.M., C.M.J., J.C.R., C.F., M.I.V., E.C.-H.O., J.J., R.L.; Software: E.C.-H.O., J.J., R.L.; Formal analysis: J.C.R., C.F., P.S., S.W., M.M.N.C., E.B.A., T.J.W., D.M.R.; Investigation: J.C.R., C.F., J.J., R.L., E.C.-H.O., P.S.; Validation: E.B.A.; Resources: S.S.M., C.M.J., A.D.S., A.F.; Writing—Original Draft: D.M.R.; Writing—Review & Editing: J.C.R., C.F., E.C.-H.O., J.J., R.L., M.I.V., P.S., C.M.J., G.G.R., A.F., T.J.W., A.D.S., H.-C.H., S.S.M., E.B.A., P.Y.; Supervision: S.S.M., A.F. All authors have read and agreed to the published version of the manuscript. Institutional Review Board Statement. The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Institutional Review Board of University of Maryland, Baltimore (GCC1488,11/16/2016). Informed Consent Statement. Informed consent was obtained from all subjects involved in the study under IRB approved protocol GCC1488. Data Availability Statement. The data presented in this study are available on request from the corresponding author. The data are not publicly available due to limitations- on patient data under the IRB protocol GCC1488. Conflicts of Interest. The University of Maryland owns patents on the subject of microtentacles and microfluidic cell tethering, and two of the authors on the current manuscript (Christopher M. Jewell and Stuart S. Martin) are listed as inventors. Christopher M. Jewell, Stuart S. Martin, and Amy Fulton are employees of the VA Maryland Health Care System. The views reported in this paper do not reflect the views of the Department of Veterans Affairs or the United States Government.Attached Files
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Additional details
- PMCID
- PMC8834106
- Eprint ID
- 113310
- DOI
- 10.3390/cancers14030800
- Resolver ID
- CaltechAUTHORS:20220207-702178000
- University of Maryland School of Medicine
- Created
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2022-02-07Created from EPrint's datestamp field
- Updated
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2022-02-14Created from EPrint's last_modified field