Ngemani Obase Bekindaka, PhD
OBASE BEKINDAKA
Nominated From: University of Washington
Research Site: University of Yaoundé
Research Area: HIV
Primary Mentor: Dara Lehman, PhD
Research Project
Mapping the intrinsic and inducible viral reservoir properties in the kidney for HIV-1 cure research in non-subtype B and C HIV patients in Cameroon
Despite significant advances in combination antiretroviral therapy (cART), HIV-1 persists in latent reservoirs, presenting a major barrier to curing the infection. Traditionally, the gut and tonsils have been extensively studied as significant reservoirs for HIV-1. However, recent findings indicate that the kidney may also serve as a substantial reservoir for latent HIV-1. This proposal aims to investigate the kidney as a reservoir for HIV-1, examining both intrinsic and inducible properties of the virus within renal tissues from patients living with HIV receiving transplants from healthy donors for 3 months postsurgery. Objectives: (1)Quantify active HIV infection in tissue by identifying infection in kidney using ISH and immunofluorescence microscopy, (2) Characterize the proviral landscape of HIV-1 in kidney tissue by identifying the intact reservoir using IPDA, (3) Assess the inducibility of HIV-1 from kidney reservoirs by evaluation the capacity of kidney reservoirs to produce infectious HIV-1 upon stimulation, (4) investigate the relationship between larger kidney reservoir size and markers of kidney dysfunction. Methods: This will be an experimental study that will run for one year and a minimum of 48 participants will be recruited: constituting of 14 participants from regional hospital Buea and 34participants from Yaoundé general hospital from whom kidney biopsies, urine and blood samples will be collected to run the following analysis; in-situ PCR, Intact proviral DNA assay, qRT-PCR, and p24 protein production using p24 Single-molecular array (SIMOA) assays, and Urinalysis for Albuminuria, creatinuria, blood creatinine and glomerular filtration rate (GFR). Statistical analysis will be conducted using SPSS, GraphPad prism, Gen AlEx, MEGA 12x, Python statistical and bioinformatic software’s will be used for data analysis and all p-values that will be less than 0.05 will be considered as statistically significant. Results: The active HIV infection in tissues will be quantified by identifying infection in kidney using in-situ PCR and ISH followed by Immunofluorescence microscopy, The proviral landscape of HIV-1 in kidney tissue will be characterized by identifying the intact reservoir using Intact proviral DNA assay (IPDA), The inducibility of HIV-1 from kidney reservoirs will be assessed by evaluation of the capacity of kidney reservoirs to produce infectious HIV-1 upon stimulation through post-stimulatory measurement of HIV-1 RNA using qRT-PCR, and p24 protein production using p24 Single-molecular array (SIMOA) assays, The relationship between larger kidney reservoir size and markers of kidney dysfunction in individuals with or at risk of kidney disease will be investigated by measuring the level of GFR, proteinuria (Albuminuria) and creatinine. Conclusion: The findings will provide critical insights into the mechanisms of HIV-1 persistence and reactivation in renal tissues, supporting the development of targeted strategies for HIV-1 eradication. This research is expected to significantly advance our understanding of HIV-1 reservoirs and contribute to the global effort to achieve a functional cure for HIV-1.
Research Significance
The burden of HIV remains a significant global health challenge, with millions living with the virus and many experiencing its devastating effects. While progress has been made in prevention and treatment, disparities exist across regions, and certain populations that are affected[23]. Initially there was hope that ART would be curative, but it quickly became clear that even though ART was able to restore CD4+ T-cell counts and suppress viral loads below levels of detection, discontinuation of treatment resulted in a rapid rebound of infection through the activation of the inactive form of the virus from reservoir organs into peripheral circulation. Therefore, identifying some of these organs and mapping out the target sites within the microenvironment is key toward HIV cure and elimination[24]. This study will provide evidence-based data that will serve as tool for the development of informed decisions for the fight against HIV. It will also lead to the improvement of clinical management of HIV infected individuals. The study is in line with the Cameroon government national strategic plan of achieving the 95-95-95 goal by 2030. To eradicate HIV as a danger to public health by 2030, Cameroon is aggressively aiming to meet the global “95-95-95” targets. These goals include 95% of individuals with HIV being aware of their status, 95% of those with HIV receiving treatment, and 95% of those receiving treatment having a suppressed viral load[25]. The Study also aligns perfectly with the U.S. President’s Emergency Plan for AIDS Relief (PEPFAR) which main objective is to help partner nations create long-lasting, nation-led solutions to the worldwide HIV/AIDS crisis. Increasing access to preventive, care, and treatment services, fortifying health systems, and coordinating HIV/AIDS programs with larger health and development projects[26]. 1.
Publications
Mentors
