Defining function of wild-type and three patient specific TP53 mutations in a zebrafish model of embryonal rhabdomyosarcoma
- The University of Texas Health Science Center at San Antonio, United States
- Hospital for Sick Children, Canada
- University of Washington, United States
Abstract
In embryonal rhabdomyosarcoma (ERMS) and generally in sarcomas, the role of wild-type and loss or gain-of-function TP53 mutations remains largely undefined. Eliminating mutant or restoring wild-type p53 is challenging; nevertheless, understanding p53 variant effects on tumorigenesis remains central to realizing better treatment outcomes. In ERMS, >70% of patients retain wild-type TP53, yet mutations when present are associated with worse prognosis. Employing a kRASG12D-driven ERMS tumor model and tp53 null (tp53-/-) zebrafish, we define wild-type and patient-specific TP53 mutant effects on tumorigenesis. We demonstrate that tp53 is a major suppressor of tumorigenesis, where tp53 loss expands tumor initiation from <35% to >97% of animals. Characterizing three patient-specific alleles reveals that TP53C176F partially retains wild-type p53 apoptotic activity that can be exploited, whereas TP53P153D and TP53Y220C encode two structurally related proteins with gain-of-function effects that predispose to head musculature ERMS. TP53P153D unexpectedly also predisposes to hedgehog expressing medulloblastomas in the kRASG12D-driven ERMS-model.
Data availability
Data sets were submitted to DRYAD, available here: doi.org/10.5061/dryad.zgmsbccb6
-
TP53 Zebrafish DataNCBI Gene Expression Omnibus, GSE213869.
-
TP53 Zebrafish DataDryad Digital Repository, doi:10.5061/dryad.zgmsbccb6.
Article and author information
Author details
Funding
Cancer Prevention and Research Institute of Texas (Training Award,RP 170345)
- Amanda E Lipsitt
St. Baldrick's Foundation
- David S Libich
Welch Foundation
- David S Libich
University of Texas Health Science Center at San Antonio (Greehey Graduate Fellowship in Children's Health)
- Paulomi Modi
Hyundai Hope On Wheels (Young Investigator Grant)
- Amanda E Lipsitt
Max and Minnie Tomerlin Voelcker Fund (Young Investigator Award)
- Myron S Ignatius
Max and Minnie Tomerlin Voelcker Fund (Young Investigator Award)
- David S Libich
University of Texas Health Science Center at San Antonio (Cancer T32 Fellow,T32CA148724)
- Kunal Baxi
University of Texas Health Science Center at San Antonio (Translational Science TL1 Fellow,TL1TR002647)
- Kunal Baxi
Cancer Prevention and Research Institute of Texas (Scholar Grant,RR160062)
- Myron S Ignatius
University of Texas Health Science Center at San Antonio (Greehey Graduate Fellowship in Children's Health)
- Nicole Rae Hensch
National Institutes of Health (R00CA1715184)
- Peter Houghton
- Myron S Ignatius
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Stephen C Ekker, Mayo Clinic, United States
Ethics
Animal experimentation: Animal studies were approved by the UT Health San Antonio Institutional Animal Care and Use Committee (IACUC) under protocol #20150015AR (mice) and #20170101AR (zebrafish). Zebrafish images were taken with specimens under tricaine anesthesia. Zebrafish tumor extraction was performed by administering high dose tricaine to minimize suffering.
Human subjects: Patient presenting with osteosarcoma signed a Consent to be part of a Repository, Epidemiology of Cancer in Children, Adolescents and Adults. In brief, this allowed for the storage of tissue, cataloging of medical information, and for research to be conducted from collected samples. The study's IRB number is HSC20080057H. Patient was informed of the risks and benefits. The umbrella study covering epidemiological study and patient-derived xenograft generation is IRB approved through UT Health San Antonio.
Version history
- Received: March 9, 2021
- Preprint posted: April 22, 2021 (view preprint)
- Accepted: June 1, 2023
- Accepted Manuscript published: June 2, 2023 (version 1)
- Version of Record published: July 5, 2023 (version 2)
Copyright
© 2023, Chen et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 989
- views
-
- 153
- downloads
-
- 1
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Defining function of wild-type and three patient specific TP53 mutations in a zebrafish model of embryonal rhabdomyosarcoma
eLife 12:e68221.
https://doi.org/10.7554/eLife.68221
Further reading
-
- Cancer Biology
- Cell Biology
Alterations in the function of K+ channels such as the voltage- and Ca2+-activated K+ channel of large conductance (BKCa) reportedly promote breast cancer (BC) development and progression. Underlying molecular mechanisms remain, however, elusive. Here, we provide electrophysiological evidence for a BKCa splice variant localized to the inner mitochondrial membrane of murine and human BC cells (mitoBKCa). Through a combination of genetic knockdown and knockout along with a cell permeable BKCa channel blocker, we show that mitoBKCa modulates overall cellular and mitochondrial energy production, and mediates the metabolic rewiring referred to as the ‘Warburg effect’, thereby promoting BC cell proliferation in the presence and absence of oxygen. Additionally, we detect mitoBKCa and BKCa transcripts in low or high abundance, respectively, in clinical BC specimens. Together, our results emphasize, that targeting mitoBKCa could represent a treatment strategy for selected BC patients in future.
-
- Cancer Biology
- Immunology and Inflammation
Solid tumors generally exhibit chromosome copy number variation, which is typically caused by chromosomal instability (CIN) in mitosis. The resulting aneuploidy can drive evolution and associates with poor prognosis in various cancer types as well as poor response to T-cell checkpoint blockade in melanoma. Macrophages and the SIRPα-CD47 checkpoint are understudied in such contexts. Here, CIN is induced in poorly immunogenic B16F10 mouse melanoma cells using spindle assembly checkpoint MPS1 inhibitors that generate persistent micronuclei and diverse aneuploidy while skewing macrophages toward a tumoricidal ‘M1-like’ phenotype based on markers and short-term anti-tumor studies. Mice bearing CIN-afflicted tumors with wild-type CD47 levels succumb similar to controls, but long-term survival is maximized by SIRPα blockade on adoptively transferred myeloid cells plus anti-tumor monoclonal IgG. Such cells are the initiating effector cells, and survivors make de novo anti-cancer IgG that not only promote phagocytosis of CD47-null cells but also suppress tumor growth. CIN does not affect the IgG response, but pairing CIN with maximal macrophage anti-cancer activity increases durable cures that possess a vaccination-like response against recurrence.
