A new study from Tel Aviv University proposes a new and unique AIDS treatment that could be developed into a vaccine or a one-time treatment for HIV patients. The study examined the engineering of B-type white blood cells in the patient's body to secrete anti-HIV antibodies in response to the virus. The technique developed in his laboratory uses B-white blood cells that would be genetically modified inside the patient's body to secrete neutralizing antibodies against the HIV virus. The gene editing was done with a CRISPR system. The researchers are able to engineer the B cells inside the patient's body using two viral vectors from the AAV family, one encodes the desired antibody and the second encodes the CRISPR system. When CRISPR cuts the desired site in the genome of the B cells it directs the introduction of the desired gene: the gene coding for the antibody against the HIV virus. On the basis of this study, we can hope that in the next few years we will be able to produce a drug against AIDS in this way, but also against other infectious diseases, for certain types of cancer caused by a virus, such as cervical cancer.Â
In this report, the scientists designed two different gRNA combinations targeting both CXCR4 and CCR5, in a single vector. The CRISPR-sgRNAs-Cas9 could successfully induce editing of CXCR4 and CCR5 genes in various cell lines and primary CD4+ T cells. Using HIV-1 challenge assays, they demonstrated that CXCR4-tropic or CCR5-tropic HIV-1 infections were significantly reduced in CXCR4- and CCR5-modified cells, and the modified cells exhibited a selective advantage over unmodified cells during HIV-1 infection. The off-target analysis showed that no non-specific editing was identified in all predicted sites. In addition, apoptosis assays indicated that simultaneous disruption of CXCR4 and CCR5 in primary CD4+ T cells by CRISPR-Cas9 had no obvious cytotoxic effects on cell viability.
A CRISPR screen conducted in a CD4+ T cell leukemia line has identified host factors required for HIV infection but dispensable for cellular survival. The results highlight sulfation on the HIV co-receptor CCR5 and cellular aggregation as potential targets for therapeutic intervention.
At the forefront of medicine, Gene Therapy brings you the latest research into genetic and cell-based technologies to treat disease. It also publishes Progress & Prospects reviews and News and Commentary articles, which highlight the cutting edge of the field.
BigField GEG Tech's insight:
Using gene editing technology, researchers at the Lewis Katz School of Medicine at Temple University have, for the first time, successfully excised a segment of HIV-1 DNA - the virus responsible for AIDS - from the genomes of living animals. The breakthrough, described online this month in the journal Gene Therapy, is a critical step in the development of a potentially curative strategy for HIV infection.
In this work, the authors used the CRISPR system to target HIV and suppresse HIV-1 replication. They demonstrated that many of these indels are indeed lethal for the virus, but that others lead to the emergence of replication competent viruses that are resistant to Cas9/sgRNA. This observation illustrates two opposite outcomes of Cas9/sgRNA action, i.e., inactivation of HIV-1 and acceleration of viral escape, thereby potentially limiting the use of Cas9/sgRNA in HIV-1 therapy.
In this work the scientists engineered CRISPR/Cas9 system to fuse nuclease deficient Cas9 with the VP64 transactivation domain. They used this tool to target 23 sites within the LTR promoter of HIV-1 and identified a “hotspot” for activation within the viral enhancer sequence. They detected consistent and effective activation of latent virus mediated by activator sgRNAs, whereas latency reversal agents produced variable activation responses.
This new generation of tool could represent a promising approach to a “functional cure” of HIV/AIDS.
Here, the authors show that sequential treatment with long-acting slow-effective release ART and AAV9- based delivery of CRISPR-Cas9 results in undetectable levels of virus and integrated DNA in a subset of humanized HIV-1 infected mice. This proof-of-concept study suggests that HIV-1 elimination is possible.
HIV presents one of the highest evolutionary rates ever detected and combination antiretroviral therapy is needed to overcome the plasticity of the virus population and control viral replication.
BigField GEG Tech's insight:
Novel strategies, such as CRISPR/Cas9 gRNA-based genome-editing, can permanently disrupt the HIV genome. However, HIV genome-editing may accelerate viral escape, questioning the feasibility of the approach.
Hultquist et al. report a high-throughput platform for the efficient, multiplex editing of host factors that control HIV infection in primary CD4+ T cells. Arrayed electroporation of CRISPR/Cas9 ribonucleoproteins (RNPs) permits the rapid generation of isogenic human cells with ablated candidate factors and identifies gene modifications that provide viral resistance.
BigField GEG Tech's insight:
Here, the scientists adapted this methodology to a high-throughput platform for the efficient, arrayed editing of candidate host factors. CXCR4 or CCR5 knockout cells generated with this method are resistant to HIV infection in a tropism-dependent manner, whereas knockout of LEDGF or TNPO3 results in a tropism-independent reduction in infection. CRISPR/Cas9 RNPs can furthermore edit multiple genes simultaneously, enabling studies of interactions among multiple host and viral factors. Finally, in an arrayed screen of 45 genes associated with HIV integrase, they identified several candidate dependency/restriction factors, demonstrating the power of this approach as a discovery platform. This technology should accelerate target validation for pharmaceutical and cell-based therapies to cure HIV infection.
Study used CRISPR technology to introduce HIV-resistance mutation into embryos.
BigField GEG Tech's insight:
Researchers in China have reported editing the genes of human embryos to try to make them resistant to HIV infection. Their paper which used CRISPR-editing tools in non-viable embryos that were destroyed after three days is only the second published claim of gene editing in human embryos.
In this work the scientists employed an RNA-guided CRISPR/Cas9 DNA editing system to precisely remove the entire HIV-1 genome spanning between 5′ and 3′ LTRs of integrated HIV-1 proviral DNA copies from latently infected human CD4+ T-cells.
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A new study from Tel Aviv University proposes a new and unique AIDS treatment that could be developed into a vaccine or a one-time treatment for HIV patients. The study examined the engineering of B-type white blood cells in the patient's body to secrete anti-HIV antibodies in response to the virus. The technique developed in his laboratory uses B-white blood cells that would be genetically modified inside the patient's body to secrete neutralizing antibodies against the HIV virus. The gene editing was done with a CRISPR system. The researchers are able to engineer the B cells inside the patient's body using two viral vectors from the AAV family, one encodes the desired antibody and the second encodes the CRISPR system. When CRISPR cuts the desired site in the genome of the B cells it directs the introduction of the desired gene: the gene coding for the antibody against the HIV virus. On the basis of this study, we can hope that in the next few years we will be able to produce a drug against AIDS in this way, but also against other infectious diseases, for certain types of cancer caused by a virus, such as cervical cancer.Â