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Hello HOPEians,

Please find below the action items from the RF2 meeting on June 7, 2023. If you have any questions about the meeting or action items please reach out to Dr. Nadia Roan and Dr. Lish Ndhlovu.

 

Actions:

• Valente Lab – Send XPB protein or plasmid to Paulo Augusto to use as a control

• Michael Corley- Send Valente lab samples to test the Western blot. Send lysis buffer recipe to test so that DNA, RNA, and protein can be extracted from the same sample)

• Andrew McAuley – Work with the Brazil team to optimize the XPB Western blot

• Ndhlovu Lab – create an inventory of received patient samples and create a list of how many cells will go into each test for a systematic analysis

• Susana will give a talk about spironolactone to the ACTG

• Matthew Parsons – Send nonhuman primate studies/ PK experiment plan to the group soon

• Valente Lab – Give DNA to Lish to look at the methylation state.

 

Updates:

During the RF2 meeting, we went around the table and heard updates from each site. Andrew McAuley shared updates from the last Spironolactone working group. They are working on the XPB western blot and described the best way to preserve the PBMC.

Vivian Avelino-Silva shared some progress on their studies at USP.  They will start collecting samples on Monday. They currently have 40 PLWH that are on spironolactone and will track age, gender, and time with diagnosis. They found 9 patients who currently have detectable viral loads, and 4 patients to start spironolactone so they can get before & after data. Luciferase is incorporated and will be offered as an optional procedure. They started the pilot study with healthy volunteers, measuring XPB to send to the Valente Lab.

Matt Parsons shared updates with MHRP. They are on schedule with their studies, but they are having trouble with sodium chloride solution from the local water in Thailand. They need to optimize before proceeding. They will have preliminary data from their primate study in the Fall.

Betty Mwesigwa with MUWRP in Uganda shared updates on the current political changes regarding Uganda’s new anti-gay legislation with the death penalty in some cases. They will continue their research but are reorganizing their procedures, recruitment, and language their staff use, in order to protect the participants. They plan to hold a meeting with their lawyer to get clear guidelines, as well as a meeting with national stakeholders to discuss the best way to move forward. In the meantime, they hope to enroll two new participants but need more guidance on which samples to prioritize.

Andrew Atkins and Scott Bowler from the Ndhlovu Lab shared data and sample processing flow from the NHS clinical studies, including 63 unique patient PIDs. They need to extract protein from all cells and send them to the Valente lab to test reservoir assay and XPB data while simultaneously extracting DNA and RNA for additional analyses. The PBMC samples have arrived, inventoried, and stored. The other plasma samples arrived and shipment is pending for the remaining.

Sara Gianella Weibel from UCSD shared they have limited amounts of tissue and have initial plans to work with Michael Corley.

Announcements

• Check out the Community Jam Board to help address some of our community’s concerns

• The next HOPE Collaboratory-wide meeting is July 24th, 9am PT/12pm ET.

• The next RF2 meeting is August 2nd

• Upcoming Speakers:

  • June 15     Dr. Mimi Ghosh

  • June 26     Drs. Steven Deeks & Michael Peluso

• 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.”

Hello HOPEians,

Please find below the action items from the RF2 meeting on April 5, 2023. If you have any questions about the meeting or action items please reach out to Dr. Nadia Roan and Dr. Lish Ndhlovu.

New Actions:

• Valente Lab – Send XPB protein or plasmid to Paulo Augusto to use as a control
• Michael Corley- Send Valente lab samples to test the Western blot. Send lysis buffer recipe to test so that DNA, RNA, and protein can be extracted from the same sample)
• Andrew McAuley – Work with the Brazil team to optimize the XPB Western blot
• Ndhlovu Lab – create an inventory of received patient samples and create a list of how many cells will go into each test for a systematic analysis

Old Actions:

• Susana will give a talk about spironolactone to the ACTG
• Matthew Parsons – Send nonhuman primate studies/ PK experiment plan to the group soon
• Valente Lab – Give DNA to Lish to look at the methylation state.

Updates:

Susana Valente gave an update on her meeting with Betty Poon and Gerard Lacourciere with the NIH during the Miami meeting to discuss Tat inhibitors. She shared that there are two paths to choose from. One might have more feedback with compounds in collaboration with Southern Research. Alternatively, another company would supply the Valente lab with novel compounds that can be tested to inhibit Tat-mediated transcription. A new chemist at the University of Florida-Scripps will advise when choosing compounds.

Susana also shared that there is a continued problem with dCA synthesis. Their new Chemist is good with the natural synthesis of products and is excited to help improve dCA production. Andrew McAuley from the Valente Lab looked at XPB degradation following spironolactone treatment using various cell types and conditions.  The Valente lab hypothesized that sodium fluoride within the SP-treated samples is creating abnormal results on the western blot (i.e. no control protein -GAPDH/Tubulin bands are showing up) Andrew will continue optimizing to try to overcome this problem and is currently working on a flow-based assay to measure XPB degradation. Ana Carolina and Paulo from the Brazil team showed their XPB western blot results. They are facing other problems with Western blot protocol and working on modifications to get better results. Andrew from the Valente lab will share their optimized protocol with the Brazil group to improve their western blots.

Lish received (estimated 107) samples today from the Natural History Studies cohort.  Their plan is to isolate and analyze the DNA and RNA simultaneously. They can save the protein from those same cells for downstream Westerns or ELISAs. Michael will send Valente lab samples to test by Western blot.

Is there a plan for single-cell RNAseq? Nadia shared that you could test the leftover cells RNA seq on fixed cells using a “fixed scRNAseq kit”. Here is the fixation protocol from 10X. 10X recommends 300K cells before fixation. The fixation kit costs about $25 a sample. You can freeze the cells to test later.

In the next meeting, Scott Bowler will give a complete update on new samples (pre and post-spironolactone tests). We will discuss non-suppressible viremia in ART-treated individuals. We would also like to hear highlights from the Tat degrader projects in the Valente Lab, p32 interaction with Tat, spironolactone updates from the Brazil team, and an update on the Last Gift meetings from Lish and Nadia.

Announcements

• Check out the Community Jam Board to help address some of our community’s concerns
• The HOPE Collaboratory-wide meeting is May 22nd 9am PT/12pm ET..
• The next RF2 meeting is June 7th
• Upcoming Speakers:
• • April 24     Dr. Jon Karn
  • May 1       Dr. Jeannette Tenthorey
  • May 15     Dr. Steve Yukl

• 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.”

Hello HOPEians,

Please find below the action items from the RF2 meeting on December 7, 2022. If you have any questions about the meeting or action items please reach out to Dr. Nadia Roan and Dr. Lish Ndhlovu. 

Updates:

Today we discussed the strengths and weaknesses from the SAB report. RF2 has done very well and the SAB did not really point out many weaknesses. One note they made was that “some candidate Tat inhibitors have EC50s in the low micromolar range and seem good candidates for optimization. However, this will require outside resources in order to pursue synthetic chemistry on several leads.” 

HOPE investigators will discuss with NIH program officers about potential strategies and resources to help with additional drug screening.  We plan to speak with Betty Poon and Gerard Lacourciere with the NIH during the Miami meeting to discuss tat-inhibitors. We could also discuss with Chris Lambros from NIAID, Avi Nath from the NIH, or Persilla Yang from Stanford about chemical biology, degraders, and protac viral targets.

Chris Lambros, PhD

National Institute of Allergy and Infectious Diseases (NIAID)

Telephone: 240-627-3093

Email: clambros@niaid.nih.gov

The next round of applications is due Jan 2023 to the Department of Health and Human Services that are internally reviewed.

HOPE investigators will reach out to Sara and Davey at the Last Gift cohort to go over potential clinical trials (combined dCA and spironolactone, Rapamycin, Brec, etc.). Clinical trials would need to be funded outside the UM1.  One potential funding source would be the ACTG:  Request for Applications- Small Clinical Trials Advancing HIV Remission and Cure (ACTG RFA). We will look at XPB degradation and Susana will discuss potential spironolactone trials to the ACTG. 

Ulrike is working with Francisco on Aim 3 using CRIPSR-based approaches. Melanie is working on dead Brec domains, but we don’t know if the effect is interfering or if this is long-lasting repression that changes the chromatin. Looking at Martin’s old work (Greene Lab), Fran is working on chromatin modifications at the LTR. We can deliver brec 1 with AV (RF3) using specific delivery methods to CD4 cells, nanobodies fused AV and higher concentrations. Priti is receiving a third batch of AAV to test on humanized mice. There is a challenge to get transgenes into T-cells lines. Other lab updates include Brazil getting their first enrollment of a patient using spironolactone.

Actions:

• Susana Valente – Coordinate with Betty Poon and Gerard Lacourciere with the NIH during the Miami meeting to discuss tat-inhibitors.

• We can connect VHL to certain binders, but how do we prove which components are binding and then develop a protac. We will coordinate with Niren to make them more permeable and maintain deficiency and solublability. 

• HOPE meetings with Last Gift

• We will look at XPB degradation and Susana will give a talk about spironolactone to the ACTG. 

• Denise – Send nonhuman primate studies/ PK experiment plan to the group soon

• Ana Leda – give DNA to Lish to look at the methylation state. 

Announcements:

• If you are attending the Miami Meeting and would like to join the HOPE dinner on 12/12 Please let Robert and me know. 

• The HOPE Collaboratory-wide meeting is on January 17th. We will hear from all the RFs.  Ulrike and Fran may be presenting for RF2 (12-minute presentation, 8-minute discussion).

• The next RF2 Meeting is February 1st. 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.”

Hello HOPEians,

Happy Holidays!

During the RF1 meeting on Dec 19, 2022, we discussed some weaknesses from the SAB report and heard a lecture from Helena Reyes-Gopar (Nixon Lab) at the Instituto Nacional De Medicina Genómica about “Stellarscope quantifies the expression of transposable elements at locus resolution in single cells.” To summarize, Helena shared that they created a tool to set in motion single-cell retrotranscriptomics; they are characterizing HERV transcription in single PBMCs, and stellarscope uses a scRNA-seq alignment and your annotation to provide a matrix that includes counts for your elements (e.g HERVs).

Down the line, the Nixon Lab plans to look at more than one donor, sort T cells and monocytes, and create a small list of TEs that are expressed. Michael Corley suggested looking at methylation/chromatin data from sorted cells to orthogonally validate and also distinguish self-expressed TEs from those co-transcribed with host genes. Ulrike mentioned that they are working with bulk and mapping to HERV by taking different public datasets and found CD4 signatures. ATAC data is generally less recalcitrant to mapping because they often overlap TE and uniquely mapping flanking sequences.

One of the weaknesses that was discussed was:

“The aims of RF1 were not all totally compelling. Testing for synergy of repressive factors might be helpful but probably not groundbreaking. Looking to HERVs as models for chromatin based repression may or may not pay off – they are mostly repressed by DNA methylation in mature cells, which allows for shutoff of embryonic ZNF genes. Perhaps this is suggesting a focus on DNA methylation, though. Not sure studying tat-independent virus will be helpful.”

In the next SAB meeting, we could do better explaining the big points in the presentations and describing the physiology in more detail. We could try to understand how different HERVs located in different loci are expressed differently and look for targeted M-seq and HERV classes.

In an upcoming Scientific Working Group – HERV & HIV on January 23rd from 10-11 am PT, Cedric, Ulrike, and Zichong are working on screening zinc finger proteins for silencers and Zichong already received some good results that will be discussed further in the next meeting.

Actions for the next meeting:

• Susana Valente – Schedule meeting with Cedric to go over PITCH approach for HERVs

• Roan Lab – Look for DUX4 expression in single-cell RNAseq tonsil datasets (+/- HIV)

• (In progress) Luisa Mori, Ana Leda, and Susana Valente considering writing a review/summary of all the HIV latency models

Announcements:

• The HOPE Collaboratory-Wide Meeting is on Tuesday, January 17, 2023, 12-1 pm PT

• There is a Scientific Working Group – HERV & HIV on January 23rd from 10-11 am PT

• The next RF1 standing meeting will be in February: Date TBD and we will discuss new HERV projects

• 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.”

Hello HOPEians,

Please find below the action items from the RF2 meeting on October 27, 2022. If you have any questions about the meeting or action items please reach out to Dr. Nadia Roan and Dr. Lish Ndhlovu. 

Updates:

Clinical Specimens

Lish gave an update on 26 patient samples from individuals on spironolactone.  Patients on eplerenone in the U.S. Military cohort were requested as a control; however, no PBMC was available.  50 controls were on amiloride.

Trevor is taking the lead for spironolactone clinical specimens.  Trevor Crowell and Brian will prepare the shipment inclusive of all controls. Collaboration between Denise/Lish/Susana -spironolactone PK in NHP samples (Matt in Thailand will send out the experimental plan to the working group).

Brazil cohort– Ana Carolina Soares mentioned that the study protocol still needs approval.  Spironolactone is not the only medication used, so enrollment is still ongoing.  No one is enrolled to date.

In vitro Experiments

Bulk RNA-seq on spironolactone-treated cells from Susana’s group may be useful in Nadia’s ex vivo primary single-cell RNA-seq study.  [~6-hour pre-treatment of spironolactone prior to infection].  Nadia Roan will continue to test PBMCs and tonsil samples. They are testing different concentrations of SP  and will block infection rates by 50% (not complete inhibition in order to leave some infected cells in spironolactone-treated condition). 

Ulrike gave an update on cell line models of HIV latency.  Generation of a cell line with a proviral reporter integrated into a region that is more heterochromatic (as reported in Elite Controllers).  CRISPR-based approaches will be used to generate these.  Alternatively, random infection and selection of clones where HIV integrates into heterochromatic regions.  Immortalization of primary cells with an hTERT plasmid – troubleshooting is underway.  Using primary T cell clones as a potential model, this needs to be optimized.

Luisa Mori is collecting data from patients treated with dCA ex vivo.  Susana discussed her group’s work on their dCA and spironolactone in combination experiments; J-Lat 10.6 cells.  The combination works better than each drug alone.  Different LRAs have been added to these conditions (8-hour LRA stimulation); GFP expression measured by flow cytometry was used as a readout.  Additionally, HIV mRNA (TAR, Gag-Pol, and Tat-Rev mRNA).  The results before Day 108 should be taken with caution due to technical issues. Focus should be placed on results past Day 108.

• Freeze/thaw greatly affects spironolactone efficacy and should be prevented by preparing more aliquots before initial freezing.  Breakdown products of Spironolactone do not work in HIV inhibition.

• Alina for Lish’s lab mentions that experimental time affects SP activity.  Susana’s experiment was 150 days. 

Susana also discussed Tat degrader TTX-881 plus Spironolactone combination experiments.  (all Tat degraders tested to date bind to Y26 residue on Tat).  After 12 days in culture, the combination condition inhibited HIV more than individually.  At 24 days, HIV was not as suppressed in this condition.  

Sonia Jablonski’s ongoing experiments:

1. Assessment of the impact of the tat degraders on tat degradation upon PROTAC cerblon knockdown in HEK293T cells

2. Optimization of the following pull downs: Tat ubiquitination, tat interaction with the components of the E3 ligase complex

3. Validate that the 3 tat degraders do not affect HIV integration

4. Ongoing analog synthesis of TT-22856 (Bannister’s lab)

5. TT-22856 was sent for pharmacokinetic analysis

6. Michael Corely / Alina Pang evaluate the 3 degraders in live cell imaging in tat 5A8 cells

Alina Pang shared some data from her dose-dependent inhibition of HIV reactivation after pre-incubation with 44863 experiments. The Tat degraders with preincubation for 5 days. She will test frozen samples for long-term effects. 

Actions:

• Denise – Send nonhuman primate studies/ PK experiment plan to the group soon

• Ana Leda – give DNA to Lish to look at the methylation state. 

Announcements:

• If you are attending the Miami Meeting and would like to join the HOPE dinner on 12/12 Please let Robert and me know. 

• The HOPE Collaboratory-wide meeting is on November 21st.

• The next RF2 Meeting is December 7th. 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.”

Hello HOPEians,

Please find below the action items from the RF2 meeting on August 29, 2022. If you have any questions about the meeting or action items please reach out to Dr. Nadia Roan and Dr. Lish Ndhlovu.

Updates:

• Lisa Henderson showed evidence that HIV transcription and translation can be observed in CSF samples from PLWH who are well-controlled on cART. The presence of these viral products is associated with neurocognitive impairment as measured by neuropsychiatric (NP) testing. They observed in vitro that targeting tat mRNA for degradation using ASOs is effective in HIV -relevant cell types (PBMC) and that knocking down tat can reduce HIV transcriptional activity and delay viral rebound; suggesting it is a viable candidate to induce ‘deep’ latency. Their next steps are to start ex-vivo experiments using patient-derived PBMCs to evaluate the effect on primary isolates. Also to start in vivo experiments in a humanized mouse model of HIV infection to evaluate the effect on tissue reservoirs and viral rebound (NIAID RAPIDD Program).

• Susana Valente explained that they solved the issue with Tat antibodies and shared dCA and SP combination treatment in J-Lat 10.6 cells using the western blot. For the cell lines, you need to block them with milk and BSA. There is difficulty detecting Tat in PBMCs and tissue samples. Currently, they are using humanized mice and monkey tissue samples. The Brazil team might be able to study this using human sample studies, and the Roan lab can test with tonsil samples.

Actions:

• Sara Gianella – Share ALS patient data and tissue availability with Avi Nath

  • Start a new MTA

  • Meeting with Sara Lemere, Avi Nath, Sara Weibel, and Lisa Henderson to discuss ALS & Endogenous Retrovirus reactivation in these HIV-positive participants.

• Sara Gianella – Collaborate with Avi Nath to mine data for defective proviral sequences

• Sara Gianella – Share morphine/opioid information on participant cohort with Lish Ndhlovu

Announcements:

• DUE TOMORROW: Call for nominations for the HOPE 2022 Awards (Instructions attached)

• 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.”

Hello HOPEians,

Please find below the action items from the RF2 meeting on July 6, 2022. If you have any questions about the meeting or action items please reach out to Dr. Nadia Roan and Dr. Lish Ndhlovu.

Updates:

• HIV Natural History Study collaboration / Trevor Crowell – New data from the exposure and timepoint definitions suggests that there are more participants than anticipated (~10) taking spironolactone and some with five or more timepoints. These longitudinal samples may include pre- and post-spironolactone administration.  Samples include PBMC but not tissues.  They are currently identifying control participants and will finalize the participant selection soon.  The MTA agreements for both Susana Valente (University of Florida) and Lish Ndhlovu (Weill Cornell) are undergoing final review and will be sent for signatures soon.

• Sara Gianella Weibel with the Last Gift Cohort gave updates – The Last Gift (NIH P01) was recently renewed until 2027. She also shared modified autopsy procedures (Rapid autopsy → collection biobanking → processing → assays). Their new assays focus on single cell resolution (VENI and VIDI project assays).  Shared lessons learned. They found heterogeneous localization of HIV DNA within the CNS and also in the periphery.  This variability was participant-specific; however, the higher levels of HIV DNA in the spinal cord stood out and will be investigated further.  HIV RNA localization studies are ongoing and so far do not correlate with HIV DNA localization.

• Esper Kallas & Lish Ndhlovu collaboration on tissue studies – Discussed studies for people taking spironolactone, goal is to use paired gut biopsy and peripheral blood analysis of HIV reservoir pre- and post- spironolactone treatment. First, they will look at spironolactone effects on general DNA transcription in HIV negative participants before they complete similar studies in HIV positive participants.  The IRB protocol has been submitted at USP and should have a response in approximately 30 days.  Initiation of the study can begin soon after IRB approval.

• Susana Valente shared data from Tat degraders that were synthesized (SR-4481 & SR-44863).  High concentrations of SR-4483 was used in a preliminary murine study, but titrations will need to be done due to the high toxicity.  MTAs are ongoing with Lish Ndhlovu at Weill Cornell and Melanie Ott at UCSF.  Susana’s group has tried to validate several additional ERCC3 (XPB) antibodies, since the previously used antibody is longer available. These new antibodies do not work for WB or IP.  The Valente lab is also continuing to study the development of viruses resistant to SP. She explained the analysis of virus potentially resistant to SP used alone from the single RhM study from Maurucio Martins.  New dCA production is still 2-3 months away.  Some of this dCA is earmarked for larger NHP studies.

• Lish shared links to:

  • Esper Kallas Oral Presentation for AIDS 2022, August 2, from 10:30-11:30 ET: Shake and bake: Promising strategies for HIV cure

  • Viral Infections & Inflammation Workshop 2022 Sep 8- Sep 9 2022

Actions:

• Sara Gianella – Share ALS patient data and tissue availability with Avi Nath

  • Start a new MTA

  • Meeting with Sara Lemere, Avi Nath, Sara Weibel, and Lisa Henderson to discuss ALS & Endogenous Retrovirus reactivation in these HIV positive participants.

• Sara Gianella – Collaborate with Avi Nath to mine data for defective proviral sequences

• Sara Gianella – Look for participants who are on spironolactone – inform Susana Valente and Nadia Roan

• Sara Gianella – Share morphine/opioid information on participant cohort with Lish Ndhlovu

Announcements:

• July 13 Guest Speaker: Dr. Mathias Lichterfeld

• The next meeting is on August 3, 2022 – Lisa Henderson will present on assays for measuring defective provirsus and HIV targeting ASOs

• If you are attending the AIDS 2022 in Montreal (July 29-August 2) please let Lish Ndhlovu and Sydney Norman know.

• Register for the HOPE Annual Meeting

• Call for nominations for the HOPE 2022 Awards (Instructions attached)

• 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.”

Hello HOPEians,

Please find below the action items from the RF2 meeting on June 1st, 20222. If you have any questions about the meeting or action items please reach out to Dr. Nadia Roan and Dr. Lish Ndhlovu. 

Updates:

• Dr. Lish Ndhlovu gave updates that they are continuing to try to make dCA with chemists at Weill Cornell.
• Dr. Brian Agan from IDCRP is joining HOPE. Several protocols have been approved to receive samples from the NHS and additional samples have been identified in RV537 of interest to HOPE.
• Dr. Ulrike Lange has sent plasmids of different Brec1 constructs to Dr. Priti Kumar and Dr. Melanie Ott. Dr. Ulrike Lange updated about the dBrec ChIP; there was a lot of plasmid background so it was difficult to call peaks, however, it did look like there were several potential off-target sites in the genome, they will look to see if these sites contain the Brec1 recombination motif. 
• Dr. Ana Carolina Soares gave an update that they finished writing their pilot project for spironolactone and aim to submit for IRB approval by the end of this week. They bought antibodies for western blotting for XBP so they can perform once they have samples. 

Actions:

• Dr. Ulrike Lange – Send cloned CRISPR off construct to Dr. Melanie Ott
• Dr. Ana Carolina Soares to send the reference for antibody for XBP to Dr. Susana Valente and Dr. Luisa Mori
• Dr. Denise Hsu to set up a subgroup meeting to discuss a potential PK study on spironolactone in NHPs. (Valente lab, Dr. Denise Hsu and team, Dr. Mauricio Martins, and Dr. Mike Cameron)
• Dr. Susana Valente to invite Dr. Mario Stevenson to give a talk about new myeloid cell paper
• (In progress) Approving and obtaining MTAs from Valente Lab (Dr. Ana Leda – MTA coordinator) 
  • Adding dCA to the MTA and getting Dr. Nadia Roan an MTA
• Dr. Susana Valente to coordinate sending DNA from dCA treated primary cells to Dr. Michael Corley to look at DNA methylation

Announcements: 

• Dr. Melanie Ott and Dr. Ulrike Lange will be hosting a CRISPR-off Efforts and Relevant Data subgroup meeting on Monday, June 13th, 10-11am PT. If you are interested and would like to join, the information is on the HOPE Calendar.
• Dr. Lish Ndhlovu shared the Department of Health and Human Services NIH Grant RFA
• The next meeting is on July 6th. 

All – If you have not already, please complete the HOPE Member Introduction form

Hello HOPEians,

Please find below the action items from the RF2 meeting on May 4th, 2022. If you have any questions about the meeting or action items please reach out to Nadia and Lish. 

In today’s meeting we got updates from:

• Valente Lab– Susana presented updates on her new Tat inhibitors. They are working to identify analogs with improved properties as well as continuing to study their mode of action.
• Ndhlovu Lab – Micheal presented updates on their live-cell imaging-based reactivation system. They have added the HC69.5 microglia clone as a cell line they can test in. Additionally, presented on new enzymatic methylation sequencing with a proviral capture method which showed good coverage of the proviral sequence.

Actions:

• Approving and obtaining MTAs from Valente Lab (Ana Leda – MTA coordinator) 
  • Adding dCA to the MTA and adding Nadia to Gladstone MTA
• Susana to coordinate sending DNA from dCA treated primary cells to Michael Corley to look at DNA methylation

Announcements: 

• The next meeting is on June 1st. 
• The email hope-rf2-members@gladstone.ucsf.edu can be used to reach the whole RF2 group.
• All – If you have not already, please complete the HOPE Member Introduction form
• Share announcements and achievements for the HOPE Twitter or Website by filling out the Social Media Shoutout form. 
• All – Email your photo to sydney.norman@gladstone.ucsf.edu

Hello HOPEians,

In Wednesday’s RF2 meeting, Lisa Henderson from Avi Nath’s lab gave an overview on taking compounds from the lab to the clinic at the NIH (slides attached). Susana gave brief updates on tat inhibitor screening, and Nadia shared new results from their spironolactone experiments in primary cells.

Please find below the action items from the RF2 meeting on March 2, 2022. If you have any questions about the meeting or action items please reach out to Nadia and Lish. 

Actions:

• All – If you have not already, please complete the HOPE Member Introduction form. Check out some of the introductions on Twitter so far!
• All – Send Lish your photo via either email or Google Drive (*Email your photo to sydney.norman@gladstone.ucsf.edu)

Announcements: 

• The next meeting is on April 6th.
• The email hope-rf2-members@gladstone.ucsf.edu can be used to reach the whole RF2 group.
• Please keep Sydney and Danielle in the loop when scheduling subgroup meetings
• Share announcements and achievements for the HOPE Twitter or Website by filling out the Social Media Shoutout form. “

The recordings from CROI discussed by Lish are available here:

• SESSION 2 – 2022 COMMUNITY HIV CURE RESEARCH WORKSHOP (FULL PLAYLIST): https://youtube.com/playlist?list=PLpzfC_l8Lo1dopnzs2-zdo4PPnoLdV0j_
• BROADLY NEUTRALIZING ANTIBODY TRIALS: https://youtu.be/m0s08rxDM5o
• HIV CURE RESEARCH IN LOW- & MIDDLE-INCOME COUNTRIES: https://youtu.be/Rz_YqMfMGN8
• HIV CURE ADVOCACY IN LOW- & MIDDLE-INCOME COUNTRIES: https://youtu.be/coWPL-WDG4o
• REPORT-BACK ON CURE-RELATED SCIENCE AT CROI 2022: https://youtu.be/vhP38HkRdlo