TY - JOUR
T1 - A novel splice variant of HYAL-4 drives malignant transformation and predicts outcome in patients with bladder cancer
AU - Lokeshwar, Vinata B.
AU - Morera, Daley S.
AU - Hasanali, Sarrah L.
AU - Yates, Travis J.
AU - Hupe, Marie C.
AU - Knapp, Judith
AU - Lokeshwar, Soum D.
AU - Wang, Jiaojiao
AU - Hennig, Martin J.P.
AU - Baskar, Rohitha
AU - Escudero, Diogo O.
AU - Racine, Ronny R.
AU - Dhir, Neetika
AU - Jordan, Andre R.
AU - Hoye, Kelly
AU - Azih, Ijeoma
AU - Manoharan, Murugesan
AU - Klaassen, Zachary
AU - Kavuri, Sravan
AU - Lopez, Luis E.
AU - Ghosh, Santu
AU - Lokeshwar, Bal L.
N1 - Funding Information:
This study is dedicated to the memory of Ms. Neetika Dhir who, together with Dr. Vinata Lokeshwar, first cloned HYAL-4 V1. Ms. Dhir passed away too soon on December 22, 2008. The research reported in this publication was partly supported by the NCI of the NIH, under the awards 1R01CA227277-01A1 (to V.B. Lokeshwar), 1F31 CA236437-01 (to D.S. Morera), and 1F31CA210612-01 to A.R. Jordan) and from the United States Army Medical Research and Development Command (USAMRDC) of the Department of Defense, under award number W81XWH-18-1-0277 (to V.B. Lokeshwar). The content of this article is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
Publisher Copyright:
© 2020 American Association for Cancer Research.
PY - 2020/7
Y1 - 2020/7
N2 - Purpose: Poor prognosis of patients with muscle-invasive bladder cancer that often metastasizes drives the need for discovery of molecular determinants of bladder cancer progression. Chondroitin sulfate proteoglycans, including CD44, regulate cancer progression; however, the identity of a chondroitinase (Chase) that cleaves chondroitin sulfate from proteoglycans is unknown. HYAL-4 is an understudied gene suspected to encode a Chase, with no known biological function. We evaluated HYAL-4 expression and its role in bladder cancer. Experimental Design: In clinical specimens, HYAL-4 wild-type (Wt) and V1 expression was evaluated by RT-qPCR, IHC, and/or immunoblotting; a novel assay measured Chase activity. Wt and V1 were stably expressed or silenced in normal urothelial and three bladder cancer cell lines. Transfectants were analyzed for stem cell phenotype, invasive signature and tumorigenesis, and metastasis in four xenograft models, including orthotopic bladder. Results: HYAL-4 expression, specifically a novel splice variant (V1), was elevated in bladder tumors; Wt expression was barely detectable. V1 encoded a truncated 349 amino acid protein that was secreted. In bladder cancer tissues, V1 levels associated with metastasis and cancer-specific survival with high efficacy and encoded Chase activity. V1 cleaved chondroitin-6-sulfate from CD44, increasing CD44 secretion. V1 induced stem cell phenotype, motility/invasion, and an invasive signature. CD44 knockdown abrogated these phenotypes. V1-expressing urothelial cells developed angiogenic, muscle-invasive tumors. V1-expressing bladder cancer cells formed tumors at low density and formed metastatic bladder tumors when implanted orthotopically. Conclusions: Our study discovered the first naturally-occurring eukaryotic/human Chase and connected it to disease pathology, specifically cancer. V1-Chase is a driver of malignant bladder cancer and potential predictor of outcome in patients with bladder cancer.
AB - Purpose: Poor prognosis of patients with muscle-invasive bladder cancer that often metastasizes drives the need for discovery of molecular determinants of bladder cancer progression. Chondroitin sulfate proteoglycans, including CD44, regulate cancer progression; however, the identity of a chondroitinase (Chase) that cleaves chondroitin sulfate from proteoglycans is unknown. HYAL-4 is an understudied gene suspected to encode a Chase, with no known biological function. We evaluated HYAL-4 expression and its role in bladder cancer. Experimental Design: In clinical specimens, HYAL-4 wild-type (Wt) and V1 expression was evaluated by RT-qPCR, IHC, and/or immunoblotting; a novel assay measured Chase activity. Wt and V1 were stably expressed or silenced in normal urothelial and three bladder cancer cell lines. Transfectants were analyzed for stem cell phenotype, invasive signature and tumorigenesis, and metastasis in four xenograft models, including orthotopic bladder. Results: HYAL-4 expression, specifically a novel splice variant (V1), was elevated in bladder tumors; Wt expression was barely detectable. V1 encoded a truncated 349 amino acid protein that was secreted. In bladder cancer tissues, V1 levels associated with metastasis and cancer-specific survival with high efficacy and encoded Chase activity. V1 cleaved chondroitin-6-sulfate from CD44, increasing CD44 secretion. V1 induced stem cell phenotype, motility/invasion, and an invasive signature. CD44 knockdown abrogated these phenotypes. V1-expressing urothelial cells developed angiogenic, muscle-invasive tumors. V1-expressing bladder cancer cells formed tumors at low density and formed metastatic bladder tumors when implanted orthotopically. Conclusions: Our study discovered the first naturally-occurring eukaryotic/human Chase and connected it to disease pathology, specifically cancer. V1-Chase is a driver of malignant bladder cancer and potential predictor of outcome in patients with bladder cancer.
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UR - http://www.scopus.com/inward/citedby.url?scp=85087469685&partnerID=8YFLogxK
U2 - 10.1158/1078-0432.CCR-19-2912
DO - 10.1158/1078-0432.CCR-19-2912
M3 - Article
C2 - 32094233
AN - SCOPUS:85087469685
SN - 1078-0432
VL - 26
SP - 3455
EP - 3467
JO - Clinical Cancer Research
JF - Clinical Cancer Research
IS - 13
ER -
