PhD Scientific Days 2025

Poster Session I. - A: Molecular Medicine

Interrogation of Antibacterial and Autoimmune Responses Using Gene-edited Neutrophilic Granulocytes

Előadó neve

Mr. Havasi Márk

Neptun code

APD775

Előadó munkahelye

Élettani Intézet, Semmelweis Egyetem

Előadó telefonszáma

+36302468477

Előadó e-mail címe

havasi.mark@stud.semmelweis.hu

Az előadás címe

Interrogation of Antibacterial and Autoimmune Responses Using Gene-edited Neutrophilic Granulocytes

Szerző(k) neve és munkahelye

Márk Havasi¹, Csaba Papp², Attila Gácser², Áron Pánczél¹, Attila Mócsai^1,3

1: Department of Physiology, Semmelweis University, Budapest, Hungary;
2: Institute of Biology, University of Szeged; Szeged, Hungary
3: HUN-REN-SE Inflammation Physiology Research Group, Budapest, Hungary

Bemutatás módja

Poszter

Szekció

Poster Session I. - A: Molecular Medicine

Language of the presentation

English

Preferred session

Molecular Medicine

Összefoglaló szövege

Introduction:
Although the role of several genes was thoroughly characterized in the migration and immune-complex induced activation of neutrophilic granulocytes (neutrophils), this process is considerably hampered by the unfeasibility of direct genetic manipulation of this cell type.
Aim:
We tried to circumvent this obstacle using CRISPR-editing of in vitro maintained, conditionally immortalised neutrophil progenitors (so-called HoxB8 cells), followed by the characterization of the resulting phenotypes in vivo.
Methods:
HoxB8 progenitors were genetically targeted using CRISPR/Cas and transplanted into irradiated recipient mice for in vivo neutrophilic differentiation. Neutrophil host-defence functions were tested in a Staphylococcus-induced peritonitis model and immune complex- induced inflammation was examined using the K/B×N serum transfer arthritis and reverse passive Arthus (RPA) reaction models.
Results:
Using our above approach, we successfully generated circulating neutrophils deficient for the Itgb2, Fcer1g or Cybb genes. Survival in our bacterial infection model was impaired in the absence of Itgb2 and Cybb genes but seemed unaffected by Fcer1g-disruption. On a mechanistic level we found severely reduced infiltration of the infected site by Itgb2-deficient neutrophils and higher intracellular bacterial burden in case of Cybb deficiency, suggesting delayed degradation of phagocytosed microbes. Inflammatory cytokine production was exaggerated in both genotypes with decreased survival. On the other hand, immune complex-induced inflammation both in terms of macroscopic signs and cytokine-production was mitigated in the absence of Itgb2 or Fcer1g but not in that of Cybb in both models (K/B×N and RPA) used.
Conclusion:
Based on our results, CRISPR-modified HoxB8 progenitors represent a rapid and efficient way to examine neutrophil functions at a single protein resolution, allowing the effect of targeted genetic manipulation on neutrophil functions to be examined in a living organism in various disease models, facilitating discovery of novel regulators of innate immune responses potentially differently involved in antibacterial and autoimmune processes and thus of great therapeutic importance.

Funding: EKÖP-2024-266.

University

Semmelweis University

Supervisor

Pánczél Áron, Mócsai Attila

Publication of my abstract

I give consent to the publication of my abstract on the website of the congress.

phd.section.field

before finishing undergraduate studies (TDK, MD-PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

9158

Start

17:42

End

17:48