454 Sequencing Study Reveals New Perspectives on Drug Resistance and Homology Related to Treatment of Invasive Inhibitors by New HIV Integrase

Research findings may serve as potential treatment options for future treatment of HIV patients

The latest data from two studies published at the 16th Conference on Retrovirus and Opportunistic Infections (CROI) in Montreal, Canada, demonstrates the ultra-deep sequencing of Roche's 454 life sciences company Genome Sequencer FLX system. The combination of traditional methods is expected to deepen the basic understanding of viral resistance and tropism, thereby promoting the development of new drugs for future selection of HIV patients. Effective drug screening for comprehensive treatment is the key to ensuring that patients have the best way to chronically suppress HIV and prevent disease progression to AIDS.

The results published at the Conference on Retroviruses and Opportunistic Infections (CROI) were completed in a series of joint research conducted by scientists at the University of Rome, Tor Vergata and Virco BVBA, at Virco Laboratories. The researchers analyzed and explained the results of sequencing HIV-infected samples with the Genome Sequencer FLX system. The study explored the resistance of HIV to a new drug called an integrase inhibitor and delved into viral tropism. The ultra-deep method of the 454 sequencing system is compared with the traditional sequencing method, which can significantly improve the sensitivity of detecting low-frequency virus mutations, that is, virus quasispecies.

The first study analyzed the long-term treatment of patients receiving antiretroviral therapy with integrated treatment with the integrase inhibitor raltegravir and optimized core therapy. The study performed multiple analyses simultaneously on multiple samples at various time points, including traditional Sanger genotyping, in vitro phenotyping, and ultra-deep sequencing. Results The researchers found that there were multiple viral quasispecies in patients who failed treatment, the level of which was much lower than the lower limit of routine detection, and the number of such quasispecies was much less in patients with successful treatment. The researchers speculate that the coexistence of many different strains of virus may contribute to the development of drug resistance and ultimately lead to treatment failure.

The researchers further suggest that the correlation between viral quasispecies identified at baseline levels and treatment failure may occur in a more complex pathway that exceeds the currently known primary drug-driven primary mutation selection. Professor Perno of Vergata University said that "the ability of 454 sequencing technology to identify very low levels of mutations provides a new perspective on how HIV can develop resistance to new drugs, such as integrase inhibitors," his colleague Ceccherini- Dr. Silberstein added, “This technology complements existing technologies and provides significant added value for HIV research.” In the second study, researchers analyzed isolated strains from HIV-infected patients, including Traditional phenotyping and genotyping, as well as deep sequencing analysis to understand HIV viral tropism. HIV tropism refers to the bias of which of the two co-receptors of CCR5 and CXCR4 is selected by a patient-specific HIV virus when it enters human CD4 cells for propagation. Determining whether a co-receptor for an HIV strain is CCR5, CXCR4, or a combination of the two is a key step in monitoring and treating HIV. A dual tropic virus can utilize either or both of the co-receptors to enter the cell. In this study, the researchers used 454 sequencing system V3 deep sequencing to analyze 10 randomly selected amphotropic isolates and studied their treatment of CCR5 antagonists and CXCR4 antagonist AMD3100 in human immune cells. Sensitivity.

Through these samples, the researchers found that most of the amphotropic viruses belong to the quasispecies that may use the R5 and CXCR4 receptors, rather than the quasispecies using the R5 and X4 receptors. This is how the HIV virus adapts to the human target cell CD4. A new perspective on the synergistic receptors of cells. Dr. Lieven Stuyver, Vice President of R&D at VIRCO BVBA, said, “This analysis helps to better understand the complex mechanisms of HIV co-receptor tropism.” “We believe that applying current technologies such as phenotyping and combining new technologies For example, the analysis of clinical outcome data by the ultra-deep sequencing of the Genome Sequencer FLX system may reveal these complex viral mechanisms and ultimately achieve better HIV treatment outcomes."

Christopher McLeod, president and CEO of 454 Life Sciences, said, "These studies have proven that 454 sequencing can be used for viral diseases and potential treatments." "Developing innovative treatments, such as new HIV drugs, often needs to be accompanied by appropriate Tools to properly select the right treatment for a particular patient. We believe that in the near future, the sensitivity, accuracy and speed of the 454 sequencing system will certainly help change the way HIV is monitored and treated."

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