Children's Medical Center Corporation

Dana-farber-cancer Institute Inc.

Richard Gregory

David A Williams

Michael Milsom

Dimitrios Iliopoulos

Elena Piskounova

George Q Daley

Srinivas R Viswanathan

Embodiments herein provide specially designed synthetic BCL11A-targeting microRNAs for RNA polymerase II expression, and methods of use to treat hemoglobinopathies such as sickle cell disease or thalassemia by increasing the expression levels of fetal hemoglobin levels.

Compositions and methods to treating hemoglobinopathies

The present embodiments provide for compositions and methods that regulate microRNA-binding protein-mediated miRNA biogensis; for example Lin28-mediated biogenesis of let-7; and in particular Lin28A-recruited 3' terminal uridylyl transferase (TUTase) uridylation of pre-let-7. A particular embodiment provide compositions and methods for screening for agents that inhibit TUTase-dependent Lin28A-mediated repression of let-7 miRNA.

Lin28-mediated control of let-7 biogenesis

Embodiment herein provide specially designed synthetic BCL11A-targeting microRNAs for RNA polymerase II expression, and methods of use to treat hemoglobinopathies such as sickle cell disease or thalassemia by increasing the expression levels of fetal hemoglobin levels. In particular illustrative embodiment, the present invention provides, in part, improved compositions and methods for achieving gene therapy in hematopoietic cells and hematopoietic precursor cells, including erythrocytes, erythroid progenitors, and embryonic stem cells. The invention further provides improved gene therapy methods for treating hematopoietic-related disorders.

The 3'-5&#x2032; exonuclease, Dis3l2, is responsible for the decay of uridylated pre-let-7 miRNA. Biochemical reconstitution assays revealed that 3&#x2032; oligouridylation stimulates Dis3l2 activity in vitro, and knockdown of Dis3l2 in mouse embryonic stem cells leads to the stabilization of pre-let-7 miRNA. These Dis3l2-depleted stem cells displayed elevated expression of pluripotency genes and delayed differentiation. The present disclosure establishes 3&#x2032; oligouridylation as an RNA decay signal for Dis3l2 and identifies the first physiological RNA substrate of this exonuclease.

Role for the perlman syndrome exonuclease Dis3l2 in the Lin28-let-7 pathway

The present embodiments provide for compositions and methods that regulate microRNA-binding protein-mediated miRNA biogenesis; for example Lin28-mediated biogenesis of let-7; and in particular Lin28A-recruited 3' terminal uridylyl transferase (TUTase) uridylation of pre-let-7. A particular embodiment provide compositions and methods for screening for agents that inhibit TUTase-dependent Lin28A-mediated repression of let-7 miRNA.

The present invention relates generally to methods to regulate microRNA (miRNA) biogenesis, in particular the regulation of the processing of pri-miRNA to mature miRNA by Lin-28 and/or variants such as Lin28B. In particular, the present invention relates to methods and compositions comprising at least one agent which inhibits Lin-28 function or activity and/or expression to increase the processing of pri-mRNA to mature miRNA. More specifically, one aspect of the invention is directed to treating and/or preventing cancer in a subject by administering an agent that inhibits Lin-28 activity or expression to a subject, preferably a human subject.

Growing community of inventors

Brookline, MA, United States of America

Richard Gregory