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EP-4735033-A2 - MRNA-ENCODED TAT WITH ATTENUATED CYTOTOXICITY FOR HIV AND SIV LATENCY REVERSAL

EP4735033A2EP 4735033 A2EP4735033 A2EP 4735033A2EP-4735033-A2

Abstract

The present invention relates to compositions and methods for targeted delivery of mRNA encoding an attenuated Tat protein and methods of use thereof for treating or preventing HIV and SIV. The compositions and methods are further useful for treatment of latent HIV or SIV.

Inventors

  • WEISSMAN, DREW
  • KREIDER, Edward

Assignees

  • The Trustees of the University of Pennsylvania

Dates

Publication Date
20260506
Application Date
20240628

Claims (20)

  1. 1. A composition comprising a delivery vehicle conjugated to a targeting domain, wherein the delivery vehicle comprises or encapsulates an mRNA molecule encoding a Tat protein or variant thereof, and further wherein the targeting domain specifically binds to CD4.
  2. 2. The composition of claim 1, wherein the delivery vehicle is selected from the group consisting of a lipid nanoparticle, a liposome, and a micelle.
  3. 3. The composition of claim 1, wherein the delivery vehicle is a lipid nanoparticle.
  4. 4. The composition of claim 1, wherein the mRNA molecule encoding a Tat protein is encapsulated in the lipid nanoparticle.
  5. 5. The composition of claim 1, wherein the Tat protein is an attenuated Tat protein about 86 amino acids in length.
  6. 6. The composition of claim 1, wherein the Tat protein comprises at least one mutation selected from the group consisting of V36A, Q66A, V67A, S68A, S77A, T23A, I39A, and L69A relative to wildtype Tat-86.
  7. 7. The composition of claim 1, wherein the Tat protein comprises V36A, Q66A, V67A, S68A, and S77A mutations.
  8. 8. The composition of claim 1, wherein the Tat protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: l, 5, 6, 11, and 13.
  9. 9. The composition of claim 1, wherein the Tat protein comprises the amino acid sequence of SEQ ID NO: 1.
  10. 10. The composition of claim 1, wherein the mRNA molecule comprises a nucleotide sequence at least 80% identical to the nucleotide sequence of SEQ ID NO:2.
  11. 11. The composition of claim 1, wherein the mRNA molecule comprises a nucleotide sequence of SEQ ID NO:2.
  12. 12. The composition of claim 1, wherein the targeting domain comprises an antibody or fragment thereof, an aptamer, a nucleic acid, a protein, a peptide, a glycan, a sugar, a hormone, or a small molecule.
  13. 13. The composition of claim 12, wherein the targeting domain comprises an anti-CD4 antibody or CD4-binding fragment thereof.
  14. 14. The composition of claim 13, wherein the anti-CD4 antibody or fragment thereof is selected from the group consisting of ibalizumab, A161A1, and CD4-binding fragments thereof.
  15. 15. A method of treating or preventing a human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV) in a subject, the method comprising administering to the subject the composition of any one of claims 1-14.
  16. 16. The method of claim 1 , wherein the HIV or SIV is latent in the subject.
  17. 17. The method of claim 15 or 16, wherein the method further comprises administering at least one additional therapeutic agent.
  18. 18. The method of claim 17, wherein the at least one additional therapeutic agent is selected from the group consisting of abacavir, emtricitabine, lamuvidine, tenofovir disoproxil fumarate, zidovudine, doravirine, efavirenz, etravirine, nevirapine, rilpivirine, atazanavir, darunavir, fosamprenavir, ritonavir, tipranavir, enfuvirtide, maraviroc, cabotegravir, dolutegravir, raltegravir, fostemsavir, idalizumab, lenacapavir, cobicistat, and combinations thereof.
  19. 19. A method of activating latent HIV or SIV in a subject comprising administering to the subject the composition of any one of claims 1-14.
  20. 20. A method of treating a subject with HIV or SIV comprising reactivating latent HIV or SIV in the subject and administering to the subject at least one anti-HIV therapy or drug.

Description

mRNA-encoded Tat with Attenuated Cytotoxicity for HIV and SIV Latency Reversal CROSS REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63/511,357, filed June 30, 2023, which is hereby incorporated by reference in its entirety. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH This invention was made with government support under AH 64570 and AI169633 awarded by the National Institutes of Health. The government has certain rights in the invention. REFERENCE TO A SEQUENCE LISTING SUBMITTED AS AN XML FILE The present application hereby incorporates by reference the entire contents of the XML file named “046483-6270-00WO_SequenceListing.xml” in XML format, which was created on June 28, 2024, and is 51,052 bytes in size. BACKGROUND OF THE INVENTION Strategies to induce a drug-free HIV cure remain research, clinical, and community priorities (Deeks, et al., 2021, Nature Medicine, 27:2085-2098). Viral latency remains a major obstacle in the development of HIV cure strategies. While currently available antiretroviral therapy (ART) can suppress active viral replication, viral reservoirs persist for years in treated individuals. The latent reservoir is a subset of the reservoir that is infected with HIV or SIV, i.e., harbors an integrated provirus, but does not express viral RNA or proteins. Replication competent virus can be isolated from these latently infected cells through stimulation of the host T cell ex vivo (Ho, Y.-C., et al., 2013, Cell, 155:540-551). As such, persistent HIV-1 reservoirs endure within an infected host despite effective antiretroviral therapy and give rise to virus rebound should therapy be interrupted (Kreider, E. F., and Bar, K. J., 2022, Current HIV/AIDS Reports, 19(3): 194-206; Li, J. Z., et al., 2016, AIDS, 30:343-353). A major contributor to persistent reservoirs are the latently infected resting CD4+ T cells (Cohn, L. B., et al., 2020, Cell Host & Microbe, 27:519-530; Ho, Y.-C., et al., 2013, Cell, 155:540-551). These cells do not express viral proteins and are indistinguishable from their uninfected counterparts (Abdel-Mohsen, M., et al., 2018, Science Translational Medicine, 10(437):eaar6759). By suppressing viral protein expression, latent viruses evade the host immune response, adoptive immunotherapy, and antiretroviral therapy and, thus, remain a major barrier to a drug-free HIV cure. Shock and kill eradication strategies target latently infected CD4+ T cells through delivery of latency reversing agents (LRAs) that reactivate quiescent virus and effectors that clear this newly reactivated virus( Margolis, D. M., et al., 2016, Science, 353(6297):aaf6517; Borducchi, E. N., et al., 2018, Nature, 563:360-364). Newer LRAs activate signaling pathways such as NF-kB, which results in transcription of the HIV-1 promoter as well as other host genes. These small molecule agonists suffer from nonspecific pleotropic effects (Zhao, M., et al., 2019, Pharmacological Research, 139:524-534), toxicities (Nixon, C. C., et al., 2020, Nature, 578:160-165; Philip, P. A., et al., 1993, JNCI, 85: 1812-1818; Kulkosky, J., et al., 2002, Journal of Infectious Diseases, 186: 1403-1411), and, importantly, potency limitations (Grau-Exposito, J., et al., 2019, PLoS Pathology, 15:el007991; Singh, V., et al., 2021, Current HI V/AIDS Reports, 18: 117-127). The viral protein Tat antagonizes latency by specifically activating viral transcription through interaction with a viral RNA feature called TAR. 89-100% of latently infected cells express TAR, demonstrating that lack of Tat expression is a major block in latency reversal (Yuki, S. A., et al., 2018, Science Translational Medicine, 10(430):eaap9927). Delivery of Tat to cells harboring latent virus, however, has remained a barrier to its development as a virus-specific LRA. Thus, there is a need in the art for improved compositions and methods for treating latent HIV. The present invention addresses this need. SUMMARY OF THE INVENTION In some embodiments, the invention provides a composition comprising a delivery vehicle conjugated to a targeting domain, wherein the delivery vehicle comprises or encapsulates an mRNA molecule encoding a Tat protein or variant thereof, and further wherein the targeting domain specifically binds to CD4. In some embodiments, the delivery vehicle is a lipid nanoparticle, a liposome, or a micelle. In some embodiments, the delivery vehicle is a lipid nanoparticle. In some embodiments, the mRNA molecule encoding a Tat protein is encapsulated in the lipid nanoparticle. In some embodiments, the Tat protein is an attenuated Tat protein about 86 amino acids in length. In some embodiments, the Tat protein comprises at least one mutation relative to a native or wild type Tat protein. Exemplary mutations include, but are not limited to, V36A, Q66A, V67A, S68A, S77A, T23A, I39A, and L69A relative to wildtype Tat-86. In some embodiments, the Tat protein comprises V36A, Q66A, V67A, S68A, and S