Dear HOPEians,
Thank you for attending the first HOPE Collaboratory-Wide Meeting on May 22, 2023. Below is a summary of the meeting, actions, and announcements.
RF2: Daniela Boehm – SMYD5 screen update
Today, Daniela Boehm, from the Ott Lab, shared an update on SMYD5 inhibitor screening. SMYD5 is a lysine methyltransferase and is required for HIV transcription. SMYD5 methylates histones and Tat in vivo. Activity tests with Tat peptides performed by collaborators at SGC Toronto showed SMYD5 has strong methylation activity at Tat aa38-72. They use promega MTase-Glo ™ Methyltransferase assay to determine the optimal SYMD5 and Tat peptide concentration with titrations. They are screening a library of known MTase inhibitors. Daniela is working on a virtual inhibitor screening with Andrii Kyrylchuk and Brian Shoichet from the UCSF School of Pharmacy. Andrii and Brian compared the SAM binding site in published SMYD family proteins SMYD2 and SMYD3 to the alphafold stucture of SMYD5. Comparing the SAM binding site, the cofactor-binding site showed a well-aligned structure between SMYD5 and other SMYD proteins. They also observed water in the binding site. Andrii and Brian virtually screened ~2 Billion fragments and picked ~200 fragments to screen in the SMYD5 inhibitor screen. Hit fragments will later be chemically modified to develop an inhibitor that is specific for SMYD5.
RF1: Qifan Wang- Prohibitin 1 & 2 and HIV transcriptional regulation
Qifan Wang, from the Valente Lab, shared his research on the activity of prohibition-2 in HIV-1 transcriptional regulations and reactivation. There are ongoing efforts to “block and lock” the HIV promoter. They identify novel protein regulators associated with active and latent HIV promoters while utilizing a compound from previous work to make HIV-infected sites latent. Some data showed enrichment from the chromatin affinity purification by CRISPR dCAS9 of transcriptionally active and latent HIV promoters. Quifan showed data from the putative “hit” HIV transcriptional regulators with both enriched active and latent LTR. There is overexpression and PHB2 inhibits HIV production in 293T cells. PHB2 reduces HIV LTR transactivation driven by Tat and PMA. PHB2 seems to bind directly to the HIV promoter and is displaced upon LTR activation with PMA. PHB2 expression is reduced in the HIV GFP+ populations, in HIV+ expressing Jurkat cells, and decreased after HIV infection in primary CD4+T cells. PHB2 restricts HIV reaction in Jurkat 10.6 cells. There is a knockdown of PHB1 also seems to increase HIV reactivation. Quifan shared some insight from the paper in Nature “Global landscape of HIV-human protein Complexes”. PHB2 expresses both in the cytoplasm and nucleus and shows some degree of co-localization. PHB2 binds to tat, likely via the basic and core domains
Community: Patricia Defechereux
Patricia Defechereux shared some community updates, recent Community projects, and positive reactions from the public. Here are some current HOPE community projects to check out:
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BLE animation: This HIV Cure Strategy EXPLAINED via Animation!
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Raif’s interview with London Patient: Scientists reveal what they think of HIV Cure research!
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Raif’s interview with Patricia: HIV Cure Research & PrEP | Patricia Defechereux x Raif Derrazi
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Podcast by Instituto Saudiversidade: Spotify HIV – From Chaos to Cure Podcasts
Tom Villa, one of HOPE’s Ambassadors, has been working with HIV cure clinical trials. In Positively Aware magazine there is a 4-part series featuring first-person accounts from participants in HIV cure clinical trials with Analytical Treatment Interruptions (ATI). Co-present with Karine Dubé at the ACTG Annual Meeting (June 13-16) on work with the ACTG Partner Protection Working Group to mitigate unintended HIV transmission during HIV cure-related clinical trials with ATI. Please check out the attached slides to see a list of upcoming Community events including the C2U Expo in Canada, a participants appreciation event in São Paulo, CAIR FOCUS Groups, etc.
RF3: Niren Murthy – LNP Targeting
Niren Murthy at Berkely shared some data from his research on LNP targeting. His primary objective for RF3 is to develop LNP/mRNA complexes that can deliver gene editor mRNA to T cells and HSPCs. Azide acetal is a new linker for generating acid-degradable drug delivery vehicles. LNPs made with acid-degradable lipids should disrupt endosomes efficiently and have lower tissue accumulation. There are too many Peg inhibitors in the function of LNPs. He shared data from LNPs with acid-degradable PEG chains transfect HSPCs in vitro with low toxicity and in vivo with Cre mRNA. For future work, he will examine the effects of GM-CSF and perform bone marrow transplants of transfected HSPCs. Looking at Acid-degradable cationic lipids, they found new lipids can be synthesized rapidly and cheaply via an automated robotic system (ARS). Lipids generated via automated LNP synthesis transfect primary T Cells and peptide lipids transfect the spleen. His next goals are to send LNP/luciferase mRNA to Kumar’s lab for testing in a humanized mouse model; deliver Cas9 mRNA and gRAN for CCR5 in vitro to T cells and HSPCs; and expand the peptide-lipids library.
Announcements
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Year 3 began May 1st, Please spend down your budget and send invoices
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Check out the Community Jam Board to help address some of our community’s concerns
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The next RF meetings: Please come prepared with some updates.
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RF1 June 28 (Ursula- RNA conformational propensities determine cellular activity)
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RF2 June 7 (TBD)
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RF3 June 5 (Rubens- Homing Endonucleases)
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Upcoming HOPE Guest Speakers:
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June 26 Steven Deeks, MD & Michael Peluso, MD
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Please Acknowledge HOPE & the NIH in any publications or presentations and notify the Program Manager. Here are some examples of what you can include.
“Research reported in this publication was supported by the NIAID of the National Institutes of Health under award number UM1AI164559, with co-funding support from NIDA, NIMH, NHLBI, the NIDDK, and the NINDS. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.”
“This research was supported by NIAID award number UM1AI164559, co-funded by NHLBI, NIDA, NIMH, NINDS, and NIDDK.”