Hello HOPEians,
Please find below the action items from the RF3 meeting on February 6, 2023. If you have any questions about the meeting or action items please reach out to Dr. Melanie Ott and Dr. Priti Kumar.
Updates:
Today we discussed the benefits of targeting CCR5 for mutagenesis and heard updates from the Kumar lab. The SAB suggested using a dual approach to help prevent future infections by targeting both the proviral genome and a host restriction factor. The caveat is that if only 30-40% of T cells have the CCR5 mutation, the viral rebound will undoubtedly occur prior to mutated cells taking over the cell milieu. Targeting efficiency has to be very high (perhaps >95%) and ideally be done in vivo; however, this is hard to achieve with nanoparticles. Up to a point, viral rebound is okay because you don’t change the size of the reservoir. There is an argument for going after the virus using something like laser ART in the most important tissue reservoirs where virus rebound can perhaps be staved off allowing time for repeated treatment with the gene editing approach. Laser ART is a nanoparticle-based ART approach that allows the slow release of ART at high concentrations in tissue reservoirs.
The Kumar lab is currently working on understanding the mutations induced by VLP-delivered CRISPR-base editors of HIV-1 provirus in Jurkat cells by sequencing the target. They are testing different guide RNAs and in the next couple of months hope to switch to mouse experiments. The lab is also adapting a base-editing approach for targeting CCR5 and CXCR4 for combination therapy.
Ott lab shared that Francisco transfects 30-40% TZMBL cells and then sorts cells with plasma DNA and not in Jurkats (TZM-bL cells are HELA cells with an integrated HIV-LTR luciferase reporter gene).
In the next RF3 meeting, we will hear from Fran and Ulrike about Brec1 delivery and what models they have used, and base editing
Here are some papers shared in today’s meeting:
- Functional T cells are capable of supernumerary cell division and longevity
- Methods and compositions for RNA-guided treatment of HIV infection
Actions:
- Kumar Lab – Send the Ott lab the mutated sequences obtained after base editing so Ursula Schulze-Gahmen can test for TAR structures
- Kumar lab – Test if you can use dead Brec 1 for base editing
- Ott and Kumar lab – Send Ulrike the constructs to test. Schedule a call to test this
- Ulrike/Fran- send Kumar lab the constructs (dead Brec1 with fusion)
- Niren – Combine and put a PEG coding on the PLGA, use conjugation coding, and do this at the beginning. Try more extensive pegylation
- Kumar lab- Connect with Ursula for biochemistry for host gene targeting
- Bring Esper onto RF3 to address their global solutions concerns
- Fran – send paper
- Michael – Share mice tissue flash freezing protocols with Priti
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- Priti can test materials sent
- Niren – Send LNPs to Priti for the Kumar lab to test – Transfection of primary human t cells with aptamer target LNPs with heart tissues
- Ulrike meeting with Fran to look at data from a previous student for Brec off suppression
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- Ulrike-Docs inducible vectors, prepping DNA, send maps and constructs to Fran
- Send new AV to Priti
- Lish – work on MTA for this (Brec, latency models), Ulrike to send MTA contact officer to Lish
Announcements:
- CRISPR for Cure – Webinar Friday, February 10th, 12-1pm PT / 3pm ET
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- Join Zoom Meeting https://temple.zoom.us/j/94549370930
- Check out the Community Jam Board to help address some of our community’s concerns
- The HOPE Collaboratory-wide meeting is on March 27.
- The next RF3 Meeting is on April 3rd. Please come prepared with some updates.
- 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.”