PhD Scientific Days 2026

Poster Session 3.U - Molecular Medicine

High-througput Identification of Factors Defining Myeloid Progenitor Survival and Function

Előadó neve

Dr. Havasi, Márk

Neptune code

APD775

Előadó munkahelye

Department of Physiology, Semmelweis University, Budapest, Hungary

Előadó telefonszáma

+36302468477

Előadó e-mail címe

havasi.mark@semmelweis.hu

Az előadás címe

High-througput Identification of Factors Defining Myeloid Progenitor Survival and Function

Szerző(k) neve és munkahelye

Márk Havasi¹, Áron Pánczél¹, Attila Mócsai^1,2,3

1: Department of Physiology, Semmelweis University, Budapest, Hungary
2: HUN-REN-SE Inflammatory Physiology Research Group, Budapest, Hungary
3: MTA-SE Inflammation Physiology Research Group, Budapest, Hungary

Bemutatás módja

Poszter

Szekció

Poster Session 3.U - Molecular Medicine

Language of the presentation

English

Preferred session

Molecular Medicine

Összefoglaló szövege

Although several genes have already been thoroughly characterized in neutrophil function, the short lifespan of these cells makes their direct manipulation unfeasible thus posing a significant obstacle in the way of neutrophil studies at single-protein resolution.
Here, we harness the ex vivo genetically tractable HoxB8 neutrophil progenitor cells that produce mature neutrophils indistinguishable from those arising from normal granulopoiesis upon in vivo adoptive transfer, to conduct high-throughput genetic screening for CRISPR-perturbations affecting proliferation/survival and in vivo differentiation phenotypes in the granulopoietic compartment.
First, we generated HoxB8 cells from primary bone marrow cells already expressing Cas9. Next, we performed a pooled CRISPR screen using the lentiviral Brie sgRNA library to enable genome-wide interrogation of gene function in HoxB8 progenitor cells. Validation of the efficiency of the library transduction confirmed adequate guide representation and uniform sgRNA distribution. Next, in vitro maintenance of the cells selected for genes required for basic cellular functions as well as myeloid progenitor specific survival signalling. As expected, top hits involved key ribosomal and chromatin proteins as well as the tyrosine kinase cKit essential for HoxB8 cell proliferation. Transduced progenitor cells were subsequently adoptively transferred into mice, where they could undergo differentiation into mature neutrophils. After maturation, cells were isolated from bone marrow and spleen for analysis. Complement-defence, MHC I function as well as cytoskeletal/migratory machinery and surprisingly also N-linked protein glycosylation and mitochondrial function related genes emerged as indispensable under in vivo circumstances compared to the in vitro maintenance.
In conclusion, lentiviral transduction of HoxB8 progenitor cells is a powerful tool for the high throughput genetic manipulation of the neutrophil compartment that can aid the unbiased identification of previously unknown single genes as well as pathways implicated in important immunological functions.

University

Semmelweis University

Supervisor

Dr. Attila Mócsai

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

phd.section.field

in doctoral studies before complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

9158

Start

14:36

End

14:39