PhD Scientific Days 2025

Pathological and Oncological Sciences I.

Development of a Human Neuroblastoma Organoid Model

Előadó neve

Mr. Tusnády Simon

Neptun code

TXLCJM

Előadó munkahelye

HUN-REN TTK, Molekuláris Élettudományi Intézet

Előadó telefonszáma

+36705177633

Előadó e-mail címe

tusy1248@gmail.com

Az előadás címe

Development of a Human Neuroblastoma Organoid Model

Szerző(k) neve és munkahelye

Simon Tusnády¹

1: HUN-REN TTK, Molekuláris Élettudományi Intézet

Bemutatás módja

Szóbeli

Szekció

Pathological and Oncological Sciences I.

Language of the presentation

Hungarian

Preferred session

Pathological and Oncological Sciences

Összefoglaló szövege

Introduction: Neuroblastoma is the most common extracranial solid tumor in children. Nearly 20% of neuroblastomas are high-risk, characterized by the amplification of the MYCN oncogene. It is known that neuroblastomas arise from sympathoblast cells of neural crest origin. However, currently no three-dimensional, replicable preclinical human neuroblastoma model is available with a uniform genetic background.

Aims: The aim of our present experiments is to develop and characterize an induced pluripotent stem cell (hiPSC)-based human neuroblastoma organoid model.

Methods: We developed a differentiation protocol to generate human neural crest organoids from hiPSCs. On days 10, 16 and 21 of differentiation neural crest markers (PHOX2B, TH, GAP43) detected by fluorescent microscopy and Western Blot were used to characterize the cell types present in the organoids. We generated transgenic iPSCs containing the MYCN gene (MYCN-GFP hiPSCs) and control iPSCs lacking MYCN (GFP hiPSCs). We verified the success of genomic integration by PCR and differentiated organoids from the transgenic hiPSCs using the previously established neural crest protocol. Then organoids were transduced with an adeno-associated virus (AAV) that induced MYCN expression in a cell-specific manner, and changes in organoid size and GFP expression were observed over time with flow cytometry and fluorescent microscopy.

Results: Expression of the neural crest markers (PHOX2B, TH, GAP43) was observed in neural crest organoids. PCR verification showed the entire transgene was integrated in the desired localization and correct orientation. The transgenic iPSCs expressed pluripotency markers SSEA-4, Oct3/4 and Nanog. Morphology and expression pattern of the differentiated organoids were similar to the control neural crest organoids. GFP expression was observed in organoids after AAV infection, and MYCN expression was detected by immunostaining.

Conclusions: We have created a hiPSC-based three-dimensional human neuroblastoma organoid model that includes tumor cells as well as healthy cells of neural crest origin forming the tumor environment. The model could be used in the future to test chemotherapeutic drugs and to study chemoresistance mechanisms in neuroblastoma.

University

Semmelweis University

Supervisor

Dr. Kornélia Szebényi

Publication of my abstract

I do not 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

szóbeli

Előadás fájl jóváhagyás

nem hagyta jóvá

Előadó

9039

Start

14:45

End

15:00