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  • An unbiased lncRNAs dropout CRISPR-Cas9 screen reveals RP11-350G8.5 as a novel therapeutic target for Multiple Myeloma
08/2024 - Blood

An unbiased lncRNAs dropout CRISPR-Cas9 screen reveals RP11-350G8.5 as a novel therapeutic target for Multiple Myeloma

Autori:

  • Grillone K.,
  • Ascrizzi S.,
  • Cremaschi P.,
  • Amato J.,
  • Polerà N.,
  • Croci O.,
  • Rocca R.,
  • Riillo C.,
  • Conforti F.,
  • Graziano R.,
  • Brancaccio D.,
  • Alcaro S.,
  • Pagano B.,
  • Randazzo A.,
  • Tagliaferri P.,
  • Iorio F.,
  • Tassone P.

Sommario:

  • We unveiled 8 lncRNAs essential for Multiple Myeloma (MM) cell fitness and associated with poor prognosis and high expression in MM patients
  • We identified lncRNA RP11-350G8.5 as a therapeutic target for MM and characterised its oncogenic role, molecular and structural features
Multiple Myeloma (MM) is an incurable malignancy characterised by altered expression of coding and non-coding genes promoting tumour growth and drug resistance. Although the crucial role of long non-coding RNAs (lncRNAs) in MM is clearly established, the function of the non-coding RNAome, which might allow the design of novel therapeutics, is largely unknown. We performed an unbiased CRISPR-Cas9 loss-of-function screen of 671 lncRNAs in MM cells and their Bortezomib (BZB)-resistant derivative. To rank functionally and clinically relevant candidates, we designed and used a bioinformatic prioritisation pipeline combining functional data from cellular screens with prognostic and transcriptional data from MM patients. With this approach, we unveiled and prioritised 8 onco-lncRNAs essential for MM cell fitness, associated with high expression and poor prognosis in MM patients. The previously uncharacterised RP11-350G8.5 emerged as the most promising target, irrespective of BZB resistance. We i) demonstrated the anti-tumoral effect obtained by RP11-350G8.5 inhibition in vitro and in vivo; ii) highlighted a modulation of the unfolded protein response and the induction of immunogenic cell death triggered by the RP11-350G8.5 knock-out, via RNA-sequencing and molecular studies; iii) characterised its cytoplasmic homing through RNA-FISH; iv) predicted its 2D structure and identified 2 G-quadruplex and 3 hairpin-forming regions by biophysical assays, including Thioflavin T, 1H-NMR and circular dichroism to pave the way to the development of novel targeted therapeutics. Overall we provided innovative insights about unexplored lncRNAs in MM and identified RP11-350G8.5 as an oncogenic target for treatment-naïve and BZB-resistant MM patients.

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