PhD Scientific Days 2024

Poster Session F - Molecular Medicine 3.

Regulation of cellular ROS levels in cutaneous melanoma by MITF

Előadó neve

Lakatos, Alexandra, MSc

Neptun code

Q6A6I1

Előadó munkahelye

Department of Physiology

Előadó telefonszáma

+36305514464

Előadó e-mail címe

lakatos.alexandra@phd.semmelweis.hu

Az előadás címe

Regulation of cellular ROS levels in cutaneous melanoma by MITF

Szerző(k) neve és munkahelye

Alexandra Lakatos¹, Dorottya Pál², Anna Sára Lengyel¹, Petra Péter¹, Botond Szabolcs¹, Elmar Lutz¹, Elisabeth M. Roider³, Akinori Kawakami³, Alicia McConnell⁴, Leonard Zon⁴, David E. Fisher⁵, Lajos V. Kemény⁶

1: HCEMM-SU Translational Dermatology Research Group, Tűzoltó street 37-47. 1094 Budapest, Hungary , Department of Physiology, Semmelweis University, Tűzoltó street 37-47. 1094 Budapest, Hungary , Department of Dermatology, Venerology and Dermatooncology, Semmelweis University, Mária street 41. 1085 Budapest, Hungary
2: HCEMM-SU Translational Dermatology Research Group, Tűzoltó street 37-47. 1094 Budapest, Hungary, Department of Physiology, Semmelweis University, Tűzoltó street 37-47. 1094 Budapest, Hungary, Department of Dermatology, Venerology and Dermatooncology, Semmelweis University, Mária street 41. 1085 Budapest, Hungary
3: Cutaneous Biology Research Center, Department of Dermatology and Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA
4: Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana-Farber Cancer Institute, Boston, Massachusetts and the Howard Hughes Medical Institute, Boston, USA
5: Cutaneous Biology Research Center, Department of Dermatology and Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
6: HCEMM-SU Translational Dermatology Research Group, Tűzoltó street 37-47. 1094 Budapest, Hungary , Department of Physiology, Semmelweis University, Tűzoltó street 37-47. 1094 Budapest, Hungary , Department of Dermatology, Venerology and Dermatooncology, Semmelweis University, Mária street 41. 1085 Budapest, Hungary, Cutaneous Biology Research Center, Department of Dermatology and Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA

Bemutatás módja

Poszter

Szekció

Poster Session F - Molecular Medicine 3.

Language of the presentation

Hungarian

Preferred session

Molecular Medicine

Összefoglaló szövege

Introductions: Microphthalmia-associated transcription factor (MITF) is crucial in melanocyte development, function, and melanoma pathogenesis. MITF amplification is common in melanoma and is associated with resistance to targeted therapies. The production of eumelanin is thought to shield the melanocytes and keratinocytes from UV damage by buffering the accumulation of UV-induced reactive oxygen species (ROS) during pigment production. We hypothesized that, besides controlling the synthesis of the antioxidant eumelanin, MITF might promote ROS clearance by contributing to the regulation of cellular ROS homeostasis.

Aims: We aim to demonstrate that MITF regulates a global antioxidant program that increases the survival of melanoma cell lines by protecting the cells from reactive oxygen species (ROS)-induced damage.

Methods: Pathway analyses using publically available datasets were made with DAVID and Cytoscape. Cells were transfected with a siRNA delivery system. Cytosolic ROS levels were measured by DCFDA fluorescent dye using flow cytometry. Cell viability was measured with trypan blue. Zebrafish were injected with MiniCoopR MITF overexpression plasmid. Zebrafish older than 15 days were euthanized by rapid chilling via submersion in ice water for 30 minutes. 8-oxoG was detected by mass spectrometry

Results: Pathways analyses indicate that MITF might regulate a transcriptional redox program in melanoma. Functional studies demonstrate the role of MITF and its target genes in reducing cytosolic and mitochondrial ROS. In vivo experiments using a zebrafish melanoma model showed that MITF decreases ROS-mediated DNA damage. Some MITF target genes, such as IDH1 and NNT, are regulated through direct MITF binding to canonical E-BOX sequences proximal to their promoters. Our data suggests that MITF is a major driver of the cellular antioxidant state

Conclusion: MITF directly (and indirectly) regulates multiple genes that collectively support the survival of melanoma cells from ROS

Funding: L.V.K. is a recipient of the János Bolyai Research Scholarship of the Hungarian Academy of Sciences and is supported by the Hungarian National Research, Development, and Innovation Office (OTKA FK138696) grant, a STIA-KFI grant from Semmelweis University. The project has received funding from the EU’s Horizon 2020 research and innovation program under grant agreement no. 739593

University

Semmelweis University

Supervisor

Lajos Vince Kemény MD, PhD

Publication of my abstract

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

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

7350

Start

15:00

End

15:03