The objective of the Pharmacokinetics and Pharmacodynamics (PK/PD) Services Core will be to provide centralized support for the PK and key PD analyses common to preclinical macaque studies in Project 2 and human clinical studies in Project 3, thereby ensuring Program's research integrity and comparability between the preclinical and clinical studies proposed. To this end, we will provide the following functions. (1) We will validate the quality of a griffithsin (GRFT) reference standard. A series of biochemical, biophysical and virological assays will be used to validate the quality of GRFT. (2) We will develop an immunoassay to validate the potency of GLP/GMP-manufactured GRFT. Upon validation of the assay at OCRP, its standard operating procedure will be transferred to Project 1 and Core B. (3) We will detect and quantify GRFT active pharmaceutical ingredient (API) and GRFT-binding antibodies in biological fluids to support PK/PD analysis in Projects 2 and 3. In addition to conventional methods based on enzyme-linked immunosorbent assays, we will develop surface plasmon resonance-based analytical methods to detect low-affinity, rapidly dissociating antibodies. As part of PK assessment, we will use HIV-1 neutralization and cell-cell fusion assays to measure the anti-HIV-1 capacity of rectal fluid samples. (4) We will analyze the impact of GRFT gel treatment on the mucosal environment to support in vivo safety assessment. To this end, we will employ a systems biology approach to comprehensively reveal the changes in the mucosal environment that may occur upon rectal administration of GRFT and GRFT combination gels. The induction of innate and adaptive immune responses in the mucosal and systemic immune compartments will be assessed using proteomic techniques. We will also use highly sensitive immunohistochemistry techniques to compare the number and location of HIV target cells, and epithelial junction proteins in rectal tissue. Additionally, the structure of rectal microbiota throughout the course of GRFT and combination gel treatment will be monitored by barcoded 16S rRNA genes sequencing to reveal the effect of microbicides formulation on the taxonomic structure of the rectal microbiota.

Public Health Relevance

Pharmacokinetics and Pharmacodynamics Services Core will facilitate preclinical and clinical research by developing and providing key assays and reagents to other components of the U19 PREVENT Program Project investigating the potent anti-HIV protein griffithsin as a candidate rectal microbicide.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
5U19AI113182-04
Application #
9276580
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
2017-07-01
Budget End
2018-06-30
Support Year
4
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Louisville
Department
Type
DUNS #
057588857
City
Louisville
State
KY
Country
United States
Zip Code
40208
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 60:6518-6531
Barton, Christopher; Kouokam, J Calvin; Hurst, Harrell et al. (2016) Pharmacokinetics of the Antiviral Lectin Griffithsin Administered by Different Routes Indicates Multiple Potential Uses. Viruses 8:
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