Exploiting embryonic niche conditions to grow Wilms tumor blastema in culture

Heather M Wojcik; Harold N Lovvorn 3rd; Melinda Hollingshead; Janene Pierce; Howard Stotler; Andrew J Murphy; Suzanne Borgel; Hannah M Phelps; Hernan Correa; Alan O Perantoni

doi:10.3389/fonc.2023.1091274

Exploiting embryonic niche conditions to grow Wilms tumor blastema in culture

Front Oncol. 2023 Mar 16:13:1091274. doi: 10.3389/fonc.2023.1091274. eCollection 2023.

Affiliations

¹ Cancer and Developmental Biology Laboratory, National Cancer Institute, Frederick, MD, United States.
² Department of Pediatric Surgery, Monroe Carell Jr. Children's Hospital at Vanderbilt University, Nashville, TN, United States.
³ Biological Testing Branch/Developmental Therapeutics Program, National Cancer Institute, Frederick, MD, United States.
⁴ Leidos Biomedical Research, National Cancer Institute at Frederick, Frederick, MD, United States.
⁵ Division of Pediatric Pathology, Monroe Carell Jr. Children's Hospital at Vanderbilt University, Nashville, TN, United States.

Abstract

Introduction: Wilms Tumor (WT), or nephroblastoma, is the most common pediatric kidney cancer. Most WTs display a "favorable" triphasic histology, in which the tumor is comprised of blastemal, stromal, and epithelial cell types. Blastemal predominance after neoadjuvant chemotherapy or diffuse anaplasia ("unfavorable" histology; 5-8%) portend a worse prognosis. Blastema likely provide the putative cancer stem cells (CSCs), which retain molecular and histologic features characteristic of nephron progenitor cells (NPCs), within WTs. NPCs arise in the metanephric mesenchyme (MM) and populate the cap mesenchyme (CM) in the developing kidney. WT blastemal cells, like NPCs, similarly express markers, SIX2 and CITED1. Tumor xenotransplantation is currently the only dependable method to propagate tumor tissue for research or therapeutic screening, since efforts to culture tumors in vitro as monolayers have invariably failed. Therefore, a critical need exists to propagate WT stem cells rapidly and efficiently for high-throughput, real-time drug screening.

Methods: Previously, our lab developed niche conditions that support the propagation of murine NPCs in culture. Applying similar conditions to WTs, we assessed our ability to maintain key NPC "stemness" markers, SIX2, NCAM, and YAP1, and CSC marker ALDHI in cells from five distinct untreated patient tumors.

Results: Accordingly, our culture conditions maintained the expression of these markers in cultured WT cells through multiple passages of rapidly dividing cells.

Discussion: These findings suggest that our culture conditions sustain the WT blastemal population, as previously shown for normal NPCs. As a result, we have developed new WT cell lines and a multi-passage in vitro model for studying the blastemal lineage/CSCs in WTs. Furthermore, this system supports growth of heterogeneous WT cells, upon which potential drug therapies could be tested for efficacy and resistance.

Keywords: Wilms Tumor; blastema; cancer stem cell; cell culture; metanephric mesenchyme.

Grants and funding

This research was supported in part by the Intramural Research Program of the NIH, NCI, Cancer and Developmental Biology Laboratory. The Translational Pathology Shared Resource at Vanderbilt University is supported by NCI/NIH Cancer Center Support Grant P30CA068485. This work was supported in part by NCI grants 4R00CA135695-03 (HNL), 5T32CA106183-06A1 (AM), and by the Section of Surgical Sciences and the Ingram Cancer Center of the Vanderbilt University Medical Center. Additional funding was obtained from the National Center for Advancing Translational Sciences [CTSA grant number UL1 TR002243 (HL)]. This project has been funded in part with federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E.