The oncolytic virus H101 combined with GNAQ siRNA‐mediated knockdown reduces uveal melanoma cell viability

Abstract

Background Uveal melanoma (UM) is a severe human malignancy with a high mortality rate, as well as high metastasis and recurrence potential. The active mutation of G protein subunit alpha q (GNAQ) or G protein subunit alpha 11 (GNA11) is a major trigger for UM. Oncolytic adenovirus H101 (H101) is the first oncolytic virus approved for clinical applications in cancer therapy by the China Food and Drug Administration. We investigated whether combining H101 with the downregulation of GNAQ expression would act synergistically in UM therapy. Methods Three UM cell lines OMM2.3 and 92.1, harboring GNAQ mutation, and OCM1, harboring B‐Raf proto‐oncogene mutation, were chosen for our research. The cellular toxicity of adenoviral infection and the cell growth rate were measured with a Cell Counting Kit‐8. Western blot analysis was used to detect GNAQ, p‐MEK1/2, YAP, and p‐YAP expression. The apoptosis and cell‐cycle distribution of cells were evaluated with annexin‐V and propidium iodide staining. Results Our results revealed that OMM2.3 and 92.1 cells were more sensitive to H101 infection than OCM1 cells. GNAQ expression was markedly reduced by small interfering RNA, siGNAQ. Combined treatment of siGNAQ and H101 inhibited the proliferation and activated the apoptosis of OMM2.3 and 92.1 cells by blocking the phosphorylation of MEK1/2 and increasing the phosphorylation of YAP. Conclusions In summary, a therapy combining H101 and siGNAQ is feasible, with potential utility as a novel targeted molecular therapy for UM, especially those carrying a GNAQ mutation.

Oncolytic Virotherapy: From Bench to Bedside

“…GNAQ mutations are known to be associated with uveal melanoma (Jager et al, 2020). Combined therapy using si-GNAQ and H101 inhibits the proliferation of uveal melanoma cells by blocking the phosphorylation of mek1/2 and promoting the phosphorylation of yap (Li et al, 2019). Immune checkpoint blockade (ICB) aims to resolve tumor escape by reversing tumor immunosuppressive signals.…”

Molecular Insights and Emerging Strategies for Treatment of Metastatic Uveal Melanoma

“…The design of simplified analogues of FR900359 capable of effective GαQ/11 inhibition, including the small molecule YM-19, opens new perspectives for pharmaceutical development [ 247 ]. Among gene regulatory approaches, a combination therapy of oncolytic adenovirus H101 and siRNA mediating GNAQ downregulation was shown to induce UM cells apoptosis in in vivo activating UM cell apoptosis [ 248 ]. Moreover, a system for conjugating siRNAs to functionalized gold nanoparticles (AuNPs) able to recognize transcripts of mutant GNAQ mRNA was developed.…”

Targeted Therapy of Uveal Melanoma: Recent Failures and New Perspectives

“…The downregulation of GNAQ mutant expression using specific short interfering RNA (siRNA) decreased GαQ protein levels in UM cell lines, resulting in a decrease in Extracellular signal–Regulated Kinases (ERK) and AKT Serine/Threonine Kinase (AKT) signaling and in 5’ Adenosine Monophosphate-activated Protein Kinase (AMPK)-dependent autophagic cell death [29]. Other studies showed that the delivery of siRNA targeting mutated GNAQ through oncolytic viruses [30] or functionalized gold nanoparticles [31] inhibits UM cell viability and growth and may be useful for future gene regulatory therapeutic approaches.…”