PMID- 35468745 OWN - NLM STAT- MEDLINE DCOM- 20220427 LR - 20220716 IS - 1471-2407 (Electronic) IS - 1471-2407 (Linking) VI - 22 IP - 1 DP - 2022 Apr 25 TI - Innovative mouse models for the tumor suppressor activity of Protocadherin-10 isoforms. PG - 451 LID - 10.1186/s12885-022-09381-y [doi] LID - 451 AB - BACKGROUND: Nonclustered mouse protocadherin genes (Pcdh) encode proteins with a typical single ectodomain and a cytoplasmic domain with conserved motifs completely different from those of classic cadherins. Alternative splice isoforms differ in the size of these cytoplasmic domains. In view of the compelling evidence for gene silencing of protocadherins in human tumors, we started investigations on Pcdh functions in mouse cancer models. METHODS: For Pcdh10, we generated two mouse lines: one with floxed exon 1, leading to complete Pcdh10 ablation upon Cre action, and one with floxed exons 2 and 3, leading to ablation of only the long isoforms of Pcdh10. In a mouse medulloblastoma model, we used GFAP-Cre action to locally ablate Pcdh10 in combination with Trp53 and Rb1 ablation. From auricular tumors, that also arose, we obtained tumor-derived cell lines, which were analyzed for malignancy in vitro and in vivo. By lentiviral transduction, we re-expressed Pcdh10 cDNAs. RNA-Seq analyses were performed on these cell families. RESULTS: Surprisingly, not only medulloblastomas were generated in our model but also tumors of tagged auricles (pinnae). For both tumor types, ablation of either all or only long isoforms of Pcdh10 aggravated the disease. We argued that the perichondrial stem cell compartment is at the origin of the pinnal tumors. Immunohistochemical analysis of these tumors revealed different subtypes. We obtained several pinnal-tumor derived (PTD) cell lines and analyzed these for anchorage-independent growth, invasion into collagen matrices, tumorigenicity in athymic mice. Re-expression of either the short or a long isoform of Pcdh10 in two PTD lines counteracted malignancy in all assays. RNA-Seq analyses of these two PTD lines and their respective Pcdh10-rescued cell lines allowed to identify many interesting differentially expressed genes, which were largely different in the two cell families. CONCLUSIONS: A new mouse model was generated allowing for the first time to examine the remarkable tumor suppression activity of protocadherin-10 in vivo. Despite lacking several conserved motifs, the short isoform of Pcdh10 was fully active as tumor suppressor. Our model contributes to scrutinizing the complex molecular mechanisms of tumor initiation and progression upon PCDH10 silencing in many human cancers. CI - (c) 2022. The Author(s). FAU - Kleinberger, Irene AU - Kleinberger I AD - Department of Biomedical Molecular Biology, Ghent University, Technologiepark-Zwijnaarde 71, 9052, Ghent, Belgium. AD - VIB-UGent Center for Inflammation Research (IRC), VIB, 9052, Ghent, Belgium. FAU - Sanders, Ellen AU - Sanders E AD - Department of Biomedical Molecular Biology, Ghent University, Technologiepark-Zwijnaarde 71, 9052, Ghent, Belgium. AD - VIB-UGent Center for Inflammation Research (IRC), VIB, 9052, Ghent, Belgium. FAU - Staes, Katrien AU - Staes K AD - Department of Biomedical Molecular Biology, Ghent University, Technologiepark-Zwijnaarde 71, 9052, Ghent, Belgium. AD - VIB-UGent Center for Inflammation Research (IRC), VIB, 9052, Ghent, Belgium. FAU - Van Troys, Marleen AU - Van Troys M AD - Department of Biomolecular Medicine, Faculty of Medicine and Health Sciences, Ghent University, 9052, Ghent, Belgium. FAU - Hirano, Shinji AU - Hirano S AD - Department of Cell Biology, Kansai Medical University, Hirakata City, Osaka, 573-1010, Japan. FAU - Hochepied, Tino AU - Hochepied T AD - Department of Biomedical Molecular Biology, Ghent University, Technologiepark-Zwijnaarde 71, 9052, Ghent, Belgium. AD - VIB-UGent Center for Inflammation Research (IRC), VIB, 9052, Ghent, Belgium. FAU - Lemeire, Kelly AU - Lemeire K AD - Department of Biomedical Molecular Biology, Ghent University, Technologiepark-Zwijnaarde 71, 9052, Ghent, Belgium. AD - VIB-UGent Center for Inflammation Research (IRC), VIB, 9052, Ghent, Belgium. FAU - Martens, Liesbet AU - Martens L AD - Department of Biomedical Molecular Biology, Ghent University, Technologiepark-Zwijnaarde 71, 9052, Ghent, Belgium. AD - VIB-UGent Center for Inflammation Research (IRC), VIB, 9052, Ghent, Belgium. FAU - Ampe, Christophe AU - Ampe C AD - Department of Biomolecular Medicine, Faculty of Medicine and Health Sciences, Ghent University, 9052, Ghent, Belgium. FAU - van Roy, Frans AU - van Roy F AD - Department of Biomedical Molecular Biology, Ghent University, Technologiepark-Zwijnaarde 71, 9052, Ghent, Belgium. frans.vanroy@ugent.be. AD - VIB-UGent Center for Inflammation Research (IRC), VIB, 9052, Ghent, Belgium. frans.vanroy@ugent.be. AD - Cancer Research Institute Ghent (CRIG), 9052, Ghent, Belgium. frans.vanroy@ugent.be. LA - eng PT - Journal Article DEP - 20220425 PL - England TA - BMC Cancer JT - BMC cancer JID - 100967800 RN - 0 (Protein Isoforms) RN - 0 (Protocadherins) SB - IM MH - Animals MH - Apoptosis/genetics MH - Cell Movement/genetics MH - Cell Proliferation/genetics MH - *Cerebellar Neoplasms MH - Humans MH - *Medulloblastoma/genetics MH - Mice MH - Protein Isoforms/genetics MH - Protocadherins PMC - PMC9040349 OTO - NOTNLM OT - Allografts OT - Conditional gene knockout OT - GFAP-Cre OT - Mouse auricular tumors OT - Mouse medulloblastoma model OT - Protocadherin-10 isoforms OT - RNA-Seq OT - Somatic stem cells OT - Tumor suppression OT - Tumor-derived cells COIS- The authors declare that they have no competing interests. EDAT- 2022/04/27 06:00 MHDA- 2022/04/28 06:00 PMCR- 2022/04/25 CRDT- 2022/04/26 05:06 PHST- 2021/06/09 00:00 [received] PHST- 2022/03/02 00:00 [accepted] PHST- 2022/04/26 05:06 [entrez] PHST- 2022/04/27 06:00 [pubmed] PHST- 2022/04/28 06:00 [medline] PHST- 2022/04/25 00:00 [pmc-release] AID - 10.1186/s12885-022-09381-y [pii] AID - 9381 [pii] AID - 10.1186/s12885-022-09381-y [doi] PST - epublish SO - BMC Cancer. 2022 Apr 25;22(1):451. doi: 10.1186/s12885-022-09381-y.