[R35] Elucidating mechanisms of spermatogonial stem cell competition
Ente: National Institute of General Medical Sciences
Scadenza: 2027-05-31
Importo max: 47.730 EUR
Paese: US
Descrizione
Project Summary
The broad, long-term objectives of this application are to characterize mechanisms that allow a competitive
germline stem cell (GSC) and its descendants to dominate the GSC population and cause super-Mendelian
inheritance. The proposal will determine how a GSC in the Drosophila testis remodels its niche and causes the
selective loss of WT neighbor GSCs. To accomplish this, the proposal will utilize immunofluorescence, genetics,
RNA interference, extended ex vivo live-cell imaging, transcriptomics, chromatin labelling, and innovative assays
of GSC competition and allele inheritance in F1 offspring. We will capitalize upon the powerful genetics available
in Drosophila, as well as the ability to unequivocally identify the niche, GSCs, differentiating germline cells, and
somatic stem cells (CySCs) and their lineage in the Drosophila testes. This proposal is supported by our
published results demonstrating that (1) loss of the transcription factor Chinmo in a GSC causes the ectopic
secretion of the extracellular matrix (ECM) protein Perlecan (Pcan), (2) this Pcan accumulates around the
endogenous niche resulting in an ectopic ECM termed the moat within the testis lumen; (3) the moat causes the
selective loss of WT neighbor GSCs, which no longer have strong adhesion with niche cells; (4) chinmo-/- GSCs
remain in the resculpted niche because they upregulate ECM-binding proteins. This proposal is also supported
by our unpublished results showing that Chinmo protein expression is promoted by an RNA-binding protein
(RBP) in GSCs and that a ZAD-ZNF protein likely acts as a Chinmo co-factor in GSCs. In the first goal, we will
determine whether clonal loss of the RBP that promotes Chinmo expression imparts that GSC with a competitive
advantage. We will also determine what regulates that RBP in GSCs and test whether loss of any regulators of
the RBP imparts a competitive advantage to a mutant GSC. In the second goal, we will determine whether
Chinmo and the ZAD-ZNF protein work together to repress Pcan by recruiting histone methyltransferases. We
will also determine how niche cells promote the ectopic Pcan produced by chinmo-/- GSCs. In the third goal, we
will test the role of somatic stem cells (CySCs) in GSC competition and assess whether they push out WT
neighbors GSCs. We will also use live-cell imaging to determine the types of GSC division that occur in chinmo-
/- GSCs. The studies in this proposal will increase the knowledge base about GSC competition and will foster
new avenues of research into mechanisms and possible treatments for human paternal age effect disorders
caused by competitive spermatogonial stem cells and for tumor cells which remodel their microenvironment to
benefit themselves and disadvantage WT neighboring cells.
Istituzione: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
PI: Erika A Bach
Progetto: 3R35GM156624-02S1
Settori: National Institute of General Medical Sciences
Vai al bando originale
Registrati gratis su Bandolo per trovare bandi compatibili con la tua azienda.