Use of trinucleotide repeat RNAs to treat cancer
Inventors
Peter, Marcus E. • Murmann, Andrea E.
Assignees
Publication Number
US-12281310-B2
Publication Date
2025-04-22
Expiration Date
2038-02-20
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Abstract
Disclosed are compositions and methods related to RNA interference (RNAi) and the use of RNAi active sequence for treating diseases and disorders. Particular disclosed are toxic RNAi active sequences such as siRNA and shRNA for killing cancer cells. The disclosed toxic RNAi active sequences typically include trinucleotide repeats and preferentially target the expression of multiple essential genes for cell survival and/or growth.
Core Innovation
The invention discloses compositions and methods involving RNA interference (RNAi) using toxic RNAi active sequences, such as siRNA or shRNA, containing trinucleotide repeats, particularly CAG and CUG repeats, for the purpose of killing cancer cells. These RNAi sequences are designed so that the guide strand includes repeat units which target and silence multiple essential genes required for cancer cell survival and/or growth, often through a mechanism similar to death induced by survival gene elimination (DISE).
The disclosed RNAi compositions typically comprise double-stranded RNA molecules with a passenger strand and a guide strand. The duplex is constructed so that the passenger strand has one or more 5' modified nucleotides preventing its incorporation into the RNA-induced silencing complex (RISC), thereby ensuring the guide strand with trinucleotide repeats, such as (CUG)n, is preferentially loaded to mediate RNAi. Upon introduction into cancer cells, these duplexes hybridize through their complementary trinucleotide repeats to form part of the double helix and powerfully suppress expression of many essential survival genes, leading to cell death.
The invention addresses the problem that traditional cancer therapies are often only marginally effective and fail to prevent resistance and metastasis, as tumor cells can evade drugs targeting single molecules or pathways. By employing trinucleotide repeat-containing RNAi sequences capable of simultaneously targeting a network of survival genes, the disclosed methods enable cell killing through activation of multiple death pathways. This combined gene silencing approach presents an entirely new way to achieve effective killing of cancer cells, with demonstrated efficacy both in vitro and in vivo.
Claims Coverage
The patent contains three independent method claims, each focusing on the introduction of specific double-stranded RNA molecules with defined trinucleotide repeat sequences to inhibit or kill cancer cells.
Double-stranded RNA with CAG/CUG trinucleotide repeats for cancer cell inhibition or killing
A method of inhibiting growth or killing a cancer cell by introducing a double-stranded RNA comprising: - A passenger strand with a (CAG)n trinucleotide repeat (n=3–10) - A guide strand with a (CUG)n trinucleotide repeat (n=3–10) - The (CUG)n and (CAG)n repeat sequences hybridize to form at least part of the duplex (15–30 nucleotides) - The passenger strand contains one or more modified nucleotides at the 5' terminus that prevent its loading into RISC - The target cancer cell is from specified types including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, or cells from adrenal gland, bladder, bone, bone marrow, brain, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, skin, testis, thymus, or uterus.
Double-stranded RNA targeting cancer cells lacking androgen receptor expression
A method of inhibiting growth or killing a cancer cell by introducing a double-stranded RNA comprising: - A passenger strand with a (CAG)n trinucleotide repeat (n=3–10) - A guide strand with a (CUG)n trinucleotide repeat (n=3–10) - The two repeat sequences hybridize to form at least part of the duplex (15–30 nucleotides) - The passenger strand contains one or more modified nucleotides at the 5' terminus that prevent its loading into RISC - The target cell is a cancer cell that does not express androgen receptor.
Double-stranded RNA with CAG/CUG repeats (n=6) targeting non-prostate/non-breast cancer cells
A method of inhibiting growth or killing a cancer cell by introducing a double-stranded RNA comprising: - A passenger strand with a (CAG)6 trinucleotide repeat - A guide strand with a (CUG)6 trinucleotide repeat - The two repeat sequences hybridize to form part of a 15–30 nucleotide duplex - The passenger strand contains one or more modified nucleotides at the 5' terminus preventing its loading into RISC - The target cancer cell is not a prostate cancer cell or a breast cancer cell.
These inventive features define methods for cancer cell inhibition or death via double-stranded RNA containing CAG and CUG trinucleotide repeats, with structural modifications to enhance guide strand specificity and application to particular cancer cell types.
Stated Advantages
The toxic RNAi sequences preferentially target and inhibit multiple essential genes for cancer cell survival and/or growth, making it difficult for cancer cells to develop resistance.
The described siRNA and shRNA methods achieve cell killing by activating multiple cell death pathways in cancer cells.
The toxic trinucleotide repeat RNAi sequences have been shown to kill a broad range of cancer cell types in vitro and effectively slow tumor growth in preclinical mouse models.
Administration of these siRNA constructs in vivo showed no gross toxicity to treated mice, indicating a favorable safety profile.
Documented Applications
Treatment of cell proliferative diseases and disorders, specifically cancer, including but not limited to adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and cancers of organs such as the adrenal gland, bladder, bone, bone marrow, brain, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, prostate, skin, testis, thymus, and uterus.
Use in pharmaceutical compositions containing toxic RNAi active sequences for administration to subjects with cancer.
Inhibition of cancer cell growth or killing of cancer cells in vitro and in vivo in preclinical models by delivering double-stranded RNA with CAG/CUG trinucleotide repeats.
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