The antiviral lectin Griffithsin (GRFT) has broad spectrum activity against human immunodeficiency viruses (HIV) types 1 and 2, herpes simplex virus type 2 (HSV-2) and hepatitis C virus (HCV) (1-4). Griffithsin-based topical microbicide formulations offer the potential for use as a multipurpose, non- antiretroviral based method to prevent HIV-1 and epidemiologically linked infections. The Project 2 objective is to show that GRFT containing rectal microbicide products are safe and effective in the Rhesus macaque model of rectal HIV transmission. These studies are designed to be complementary to the human clinical studies proposed in Project 3. We have three specific aims:
Aim 1 is to evaluate the safety, pharmacokinetics and pharmacodynamics (PK/PD) of GRFT carbopol gel. We will conduct a three stage placebo controlled PK/PD study in adult male Rhesus macaques with the GRFT rectal gel formulation and matched placebo gel supplied by Core B. The proposed macaque study is designed to address specific translational questions concerning immunotoxicity and health of the mucosal environment after GRFT exposure. Favorable stage 1 outcomes include detection of active HIV-1 inhibitory concentrations of GRFT in rectal fluids and acceptable clinical safety metrics, which allows progression to the efficacy Stage 2. Using a cohort of GRFT treatment experienced animals will allow us to determine whether pre-existing immunity to GRFT affects product efficacy. Stage 3 will test the GRFT and placebo products in SHIV infected animals, where we will compare amounts of virus shed in experimental versus control animals, providing an additional safety metric.
Aim 2 is to assess the impact of GRFT treatment on the mucosal immune environment. We will use multiplexed immunoassays and gene expression arrays to measure markers of inflammatory immune responses. The highly sensitive immunohistochemistry (IHC) techniques refined in Prof. Kristina Broliden's laboratory will be applied to compare the number, distribution and activation state of HIV target cells and expression of HIV receptor targets CD4, CCR5, DC-SIGN and Langerin plus markers of epithelial integrity in rectal biopsies from animals treated with GRFT active drug or matched placebo. Flow cytometry analyses will also enumerate and phenotype lymphocytes in biopsy tissues.
Aim 3 is to assess the impact of GRFT treatment on the mucosal environment in the rectum using a systems biology approach that utilizes modern mass spectrometry proteomics assays to compare tissues and rectal secretions from animals that receive active drug versus placebo, and to evaluate the effect of GRFT treatment on the rectal microbiota of treated animals.
The global incidence of HIV-1 infection is declining, but not in populations that practice unprotected anal intercourse (URAI), highlighting urgent need for attention to the preventative health needs in communities where men who have sex with men are especially stigmatized. Our rectal gel formulations have potential to prevent HIV-1 transmission via URAI, but must first be tested for safety and effectiveness in animals. Our studies aim to provide a comprehensive set of data in support of further product development, and clinical testing of the griffithsin gel product in the PREVENT clinical trial.
|Grooms, Tiffany N; Vuong, Hung R; Tyo, Kevin M et al. (2016) Griffithsin-Modified Electrospun Fibers as a Delivery Scaffold to Prevent HIV Infection. Antimicrob Agents Chemother :|
|Fuqua, Joshua L; Wanga, Valentine; Palmer, Kenneth E (2015) Improving the large scale purification of the HIV microbicide, griffithsin. BMC Biotechnol 15:12|
|Fuqua, Joshua L; Hamorsky, Krystal; Khalsa, Guruatma et al. (2015) Bulk production of the antiviral lectin griffithsin. Plant Biotechnol J 13:1160-8|