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ROR1-targeted delivery of miR-29b induces cell cycle arrest and therapeutic benefit in vivo in a CLL mouse model

Chi-Ling Chiang, Swagata Goswami, Frank W. Frissora, Zhiliang Xie, Pearlly S. Yan, Ralf Bundschuh, Logan A. Walker, Xiaomeng Huang, Rajeswaran Mani, Xiaokui M. Mo, Sivasubramanian Baskar, Christoph Rader, Mitch A. Phelps, Guido Marcucci, John C. Byrd, L. James Lee and Natarajan Muthusamy

Key Points

  • Clinically available miR-29b exerts potent antileukemic activity in primary CLL cells in vitro and in an hROR1 × TCL1 mouse model in vivo.

  • Leukemia-targeted delivery of miR-29b via ROR1 induces epigenetic reprogramming and p21-dependent cell cycle arrest in vivo.

Abstract

Chronic lymphocytic leukemia (CLL) occurs in 2 major forms: aggressive and indolent. Low miR-29b expression in aggressive CLL is associated with poor prognosis. Indiscriminate miR-29b overexpression in the B-lineage of mice causes aberrance, thus warranting the need for selective introduction of miR-29b into B-CLL cells for therapeutic benefit. The oncofetal antigen receptor tyrosine kinase orphan receptor 1 (ROR1) is expressed on malignant B-CLL cells, but not normal B cells, encouraging us with ROR1-targeted delivery for therapeutic miRs. Here, we describe targeted delivery of miR-29b to ROR1+ CLL cells leading to downregulation of DNMT1 and DNMT3A, modulation of global DNA methylation, decreased SP1, and increased p21 expression in cell lines and primary CLL cells in vitro. Furthermore, using an Eμ-TCL1 mouse model expressing human ROR1, we report the therapeutic benefit of enhanced survival via cellular reprograming by downregulation of DNMT1 and DNMT3A in vivo. Gene expression profiling of engrafted murine leukemia identified reprogramming of cell cycle regulators with decreased SP1 and increased p21 expression after targeted miR-29b treatment. This finding was confirmed by protein modulation, leading to cell cycle arrest and survival benefit in vivo. Importantly, SP1 knockdown results in p21-dependent compensation of the miR-29b effect on cell cycle arrest. These studies form a basis for leukemic cell–targeted delivery of miR-29b as a promising therapeutic approach for CLL and other ROR1+ B-cell malignancies.

  • Submitted October 31, 2018.
  • Accepted April 29, 2019.
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