Development and analysis of response biomarkers for miR-323a-5p therapy in Neuroblastoma

Abstract

Neuroblastoma (NB) is the most common embryonal cancer among children. It originates

in the peripheral sympathetic nervous system and can metastasize to various parts of

the body. Unfortunately, current treatments do not prevent relapse in high-risk patients,

suggesting the need for new therapies. RNA-based therapies, especially those involving

microRNAs (miRNAs), show potential in the treatment of NB, with miR-323a-5p being

one of the miRNAs that exhibits more potential as a tumor suppressor, making it a

promising therapeutic candidate. However, effective delivery of miR-323a-5p is

challenging due to its degradation in the bloodstream and cellular uptake. Nanoparticles

offer a potential solution and enabled the in vivo delivery of miR-323a-5p, demonstrating

efficacy in known molecular targets. Although these target genes phenotypically reflect

the effects of miR-323a-5p, they may not optimally detect this nanomedicine efficacy,

suggesting the need of working on the study of a response biomarker that is further

modulated.

In this study, by RNA-seq analysis of NB samples treated with miR-323a-5p, potential

biomarkers of response that could act as targets to determine the antitumor efficacy of

this miRNA in vivo were identified and analyzed. These results were validated by qPCR,

considering two different delivery systems: Lipofectamine 2000 (LF 2000), commonly

used in in vitro studies and Quatsomes (QS), a nanoparticle-based delivery system.

RNA-seq analysis identified 8 genes that were modulated by miR-323a-5p. Although two

of the genes could not be amplified, the RNA-seq results were largely confirmed by

qPCR. Although modulation was detected, the in vitro results do not reveal a clear

candidate response biomarker that significantly improves upon those currently used. The

study also reveals significant variation in gene modulation between delivery systems,

with QS being less efficient compared to LF 2000. These findings suggest the need for

future in vivo validation, studies using different NB cell lines and consideration of multiomics approaches to better understand the therapeutic impact of miR-323a-5p in NB