TY - JOUR
T1 - The component of the m6A writer complex VIRMA is implicated in aggressive tumor phenotype, DNA damage response and cisplatin resistance in germ cell tumors
AU - Miranda-Gonçalves, Vera
AU - Lobo, João
AU - Guimarães-Teixeira, Catarina
AU - Barros-Silva, Daniela
AU - Guimarães, Rita
AU - Cantante, Mariana
AU - Braga, Isaac
AU - Maurício, Joaquina
AU - Oing, Christoph
AU - Honecker, Friedemann
AU - Nettersheim, Daniel
AU - Looijenga, Leendert H.J.
AU - Henrique, Rui
AU - Jerónimo, Carmen
N1 - Publisher Copyright:
© 2021, The Author(s).
PY - 2021/8/25
Y1 - 2021/8/25
N2 - BACKGROUND: Germ cell tumors (GCTs) are developmental cancers, tightly linked to embryogenesis and germ cell development. The recent and expanding field of RNA modifications is being increasingly implicated in such molecular events, as well as in tumor progression and resistance to therapy, but still rarely explored in GCTs. In this work, and as a follow-up of our recent study on this topic in TGCT tissue samples, we aim to investigate the role of N6-methyladenosine (m6A), the most abundant of such modifications in mRNA, in in vitro and in vivo models representative of such tumors.METHODS: Four cell lines representative of GCTs (three testicular and one mediastinal), including an isogenic cisplatin resistant subline, were used. CRISPR/Cas9-mediated knockdown of VIRMA was established and the chorioallantoic membrane assay was used to study its phenotypic effect in vivo.RESULTS: We demonstrated the differential expression of the various m6A writers, readers and erasers in GCT cell lines representative of the major classes of these tumors, seminomas and non-seminomas, and we evidenced changes occurring upon differentiation with all-trans retinoic acid treatment. We showed differential expression also among cells sensitive and resistant to cisplatin treatment, implicating these players in acquisition of cisplatin resistant phenotype. Knockdown of VIRMA led to disruption of the remaining methyltransferase complex and decrease in m6A abundance, as well as overall reduced tumor aggressiveness (with decreased cell viability, tumor cell proliferation, migration, and invasion) and increased sensitivity to cisplatin treatment, both in vitro and confirmed in vivo. Enhanced response to cisplatin after VIRMA knockdown was related to significant increase in DNA damage (with higher γH2AX and GADD45B levels) and downregulation of XLF and MRE11.CONCLUSIONS: VIRMA has an oncogenic role in GCTs confirming our previous tissue-based study and is further involved in response to cisplatin by interfering with DNA repair. These data contribute to our better understanding of the emergence of cisplatin resistance in GCTs and support recent attempts to therapeutically target elements of the m6A writer complex.
AB - BACKGROUND: Germ cell tumors (GCTs) are developmental cancers, tightly linked to embryogenesis and germ cell development. The recent and expanding field of RNA modifications is being increasingly implicated in such molecular events, as well as in tumor progression and resistance to therapy, but still rarely explored in GCTs. In this work, and as a follow-up of our recent study on this topic in TGCT tissue samples, we aim to investigate the role of N6-methyladenosine (m6A), the most abundant of such modifications in mRNA, in in vitro and in vivo models representative of such tumors.METHODS: Four cell lines representative of GCTs (three testicular and one mediastinal), including an isogenic cisplatin resistant subline, were used. CRISPR/Cas9-mediated knockdown of VIRMA was established and the chorioallantoic membrane assay was used to study its phenotypic effect in vivo.RESULTS: We demonstrated the differential expression of the various m6A writers, readers and erasers in GCT cell lines representative of the major classes of these tumors, seminomas and non-seminomas, and we evidenced changes occurring upon differentiation with all-trans retinoic acid treatment. We showed differential expression also among cells sensitive and resistant to cisplatin treatment, implicating these players in acquisition of cisplatin resistant phenotype. Knockdown of VIRMA led to disruption of the remaining methyltransferase complex and decrease in m6A abundance, as well as overall reduced tumor aggressiveness (with decreased cell viability, tumor cell proliferation, migration, and invasion) and increased sensitivity to cisplatin treatment, both in vitro and confirmed in vivo. Enhanced response to cisplatin after VIRMA knockdown was related to significant increase in DNA damage (with higher γH2AX and GADD45B levels) and downregulation of XLF and MRE11.CONCLUSIONS: VIRMA has an oncogenic role in GCTs confirming our previous tissue-based study and is further involved in response to cisplatin by interfering with DNA repair. These data contribute to our better understanding of the emergence of cisplatin resistance in GCTs and support recent attempts to therapeutically target elements of the m6A writer complex.
KW - Adenosine/analogs & derivatives
KW - Antineoplastic Agents/therapeutic use
KW - Cell Line, Tumor
KW - Cisplatin/therapeutic use
KW - DNA Damage
KW - Drug Resistance, Neoplasm/physiology
KW - Gene Knockdown Techniques
KW - Humans
KW - Male
KW - Methyltransferases/physiology
KW - Neoplasms, Germ Cell and Embryonal/drug therapy
KW - RNA-Binding Proteins/genetics
UR - http://www.scopus.com/inward/record.url?scp=85113554533&partnerID=8YFLogxK
U2 - 10.1186/s13046-021-02072-9
DO - 10.1186/s13046-021-02072-9
M3 - Article
C2 - 34446080
SN - 0392-9078
VL - 40
SP - 268
JO - Journal of experimental & clinical cancer research : CR
JF - Journal of experimental & clinical cancer research : CR
IS - 1
ER -