Structure‐function relationships for the lncRNA SChLAP1 in aggressive prostate cancer

Abstract

Advances in deep sequencing technologies have facilitated the identification and annotation of thousands of long noncoding RNAs (lncRNAs) across the transcriptome. LncRNAs are documented to play critical housekeeping roles within the cell and are implicated in a wide variety of diseases, including cancer. While studies into lncRNA function in cancer abound, there are limited examples to date of detailed lncRNA structural analyses to enable understanding of structure‐function relationships. Understanding structure‐function relationships would increase our insight into the noncoding transcriptome and yield potential avenues for targeting lncRNAs implicated in disease. The lncRNA Second Chromosome Locus Associated with Prostate 1 (SChLAP1) has been identified in multiple clinical studies as a predictive biomarker and molecular driver of aggressive prostate cancer. While several protein interactors have been identified for SChLAP1 to date, structural insight into SChLAP1:protein recognition has not yet been explored. We believe that structural analysis of SChLAP1 will assist in designing specific therapeutic strategies to inhibit SChLAP1:protein interactions implicated in prostate cancer. To this end, we performed Selective 2′‐Hydroxyl Acylation analyzed by Primer Extension with Mutational Profiling (SHAPE‐MaP) and dimethyl sulfate (DMS)‐MaP in vitroand in cellulo. This approach yielded the first secondary structure model of SChLAP1, which revealed a complex architecture with a wide variety of secondary structures throughout the length of the transcript. Analyzing our in‐cell probing data with the ΔSHAPE algorithm, we identified protein binding regions within SChLAP1 and mapped them to non‐human primate‐conserved exons within the transcript. Finally, we identified a smaller, highly structured fragment of SChLAP1 that houses multiple putative protein binding sites implicated in prostate cancer. We believe that this region of SChLAP1 is amenable to therapeutic targeting and may be used as a smaller construct for in vitrodevelopment of SChLAP1‐targeting therapeutics. We also believe this fragment may be amenable to 3D biophysical analyses, such as X‐ray crystallography or cryo‐EM, to further enhance structural understanding of SChLAP1:protein complexation. Ongoing work is focused on determining the sufficiency of this fragment for protein recognition and characterizing potential magnesium‐dependent tertiary structures. We believe this work will facilitate the development of specific therapeutic strategies for SChLAP1 and contribute to the growing need of characterizing structure‐function relationships within lncRNAs.

LncRNA weighted gene co-expression network analysis reveals novel biomarkers related to prostate cancer metastasis

“…LncRNA mediated mechanisms have been associated with epigenetic changes in prostate cancer. According to the results of the COSMIC-021, combination of the cabozantinib, a kind of antiangiogenic drugs, with the immune checkpoint inhibitor atezolizumab achieved encouraging activity in patients with metastatic castration-resistant PC (mCRPC), while multiple previous phase III clinical trials 5,6 in refractory CRPC have demonstrated increased toxicity with no clinical benefit. Tumor angiogenesis is regulated through interaction between anti-angiogenic factors and proangiogenic within the tumor microenvironment.…”

“…Furthermore, the Progensa PCA3 test was successfully approved by the US Food and Drug Administration (FDA) in 2012 [ 5 ]. The lncRNA Second Chromosome Locus Associated with Prostate 1 (SChLAP1) has also been identified as a molecular driver and predictive biomarker of aggressive PCa in multiple clinical studies [ 6 ]. PCa is the foremost non-cutaneous malignancy in men worldwide, with increasing morbidity and mortality.…”