Durham Research Center, 8020
985880 University of Nebraska Medical Center
Omaha, NE 68198-5880
HIV-1, Humanized mice, nanoART, Pharmacodynamics, HIV-1 associated neurocognitive disorders (HAND), drug-drug interaction, detection and targeting of HIV-1 reservoirs, HIV-1 elimination, CRISPR-Cas9 based targeting
UNMC Today | February 6, 2018
Chemically modified drug shows promise for HIV treatment, elimination
My major focus is to understand the drug-drug interactions on molecular pathways for HIV therapeutics and elimination. Thirty-seven years after the discovery that HIV is the causative agent of AIDS, there is still the problem of drug adherence, drug dosing and appearance of drug resistant strains, and unfruitful targeting of drugs to the latent reservoirs in the patients. There are only two documented cases of HIV-1 functional cure till date. The subsequent success was stalled because of 1) limited therapeutic access to viral reservoirs, 2) the known rapid spread of viral infection, 3) much higher numbers of viral susceptible cells in HIV-1 target tissues and 4) an inability to completely eliminate integrated proviral DNA. Viral rebound is uniformly seen after cessation of antiretroviral therapy (ART). To address each of these limitations, I am working with a medicinal chemistry team to improve drug penetrance across cell and tissue barriers and to improve control over ongoing viral infection. We reasoned that, under conditions of optimal viral restriction, deployment of CRISPR/Cas9 proviral DNA excision strategies targeting specific host genes responsible for viral entry and multiple regions of HIV-1 genome would facilitate elimination of HIV-1 from the host genome. This will be validated using multiple highly sensitive molecular detection in vitro and in vivo assays.
- Targeting HIV-1 elimination using a sequential combinatorial treatment paradigm in Humanized Mice
- Detection and targeting of HIV-1 reservoirs using multimodal approach: In our collaborative work with Drs. Howard Gendelman and Bhavesh Kevadiya, we are trying to develop a new theranostic nanosystem to optimize site-directed antiretroviral drug (ARV) delivery to infected cells and tissues to enable maximal viral restriction in lymphoid reservoirs and brain in a novel mouse model of HIV disease and to track and treat HIV-1.
- Understanding the cytokine and metabolic changes at the cellular and molecular level in HIV-infected and/or treated and virally suppressed humanized mice tissue samples isolated from myeloid, lymphoid, and CNS compartments
- Development of ultra-sensitive in vivo detection assay system for HIV-1, studying hu-NSG mouse-based viral outgrowth assay (MVOA) that can successfully recover virus from virally suppressed humanized mouse tissues isolated from myeloid, lymphoid, and peripheral compartments. These mVOA assays will be employed in future to interrogate HIV-1 latency from individual tissue or cell type of interest and can become standard validation assays to confirm HIV-1 elimination.
- Development of a testing pharmacodynamic (PD) platform for the development of antiretroviral and adjunctive therapies for peripheral HIV-1, and for its related associated neurocognitive disorders
- Generation and HIV-1 infection drug efficacy studies using Hu-PBL mice
- Flow cytometry on blood and tissues
- Immunohistochemistry (IHC)
- Immunofluorescence (IF) microscopy and quantification using Nuance
- Harvesting of mice organs and tissue RNA and DNA isolation
- Cell culture and infections of primary cells with HIV-1
- Reverse transcriptase (RT) assay
- HIV-1 Viral load measurements from plasma and tissues
- Western blots
- Conventional PCRs, Real time-qRT PCR and droplet digital PCR, RNAscope
- Dash PK, Alomar F A, Hackfort B T, Su H, Conaway A, Poluektova LY, Gendelman H E, Gorantla S and Bidasee K R. HIV-1 Associated Left Ventricular Cardiac Dysfunction in Humanized Mice. Sci Rep. 2020 Jun 16; 10 (1) 9746.
- Su H, Sravanam S, Gorantla S, Kaminski R, Khalili K, Poluektova L, Gendelman HE, Dash PK. Amplification of Replication Competent HIV-1 by adoptive transfer of Human cells from Tissues of Infected Humanized Mice. Front Cell and Infect Micro. 2020 Feb 11; 10:38.
- Dash PK, Kevadiya B, Su H, Banoub M, Gendelman HE. Pathways towards Human Immunodeficiency Virus Elimination. EBioMedicine 2020 Feb 27;53102667.
- Dash PK, R Kaminski, R Bella, H Su, S Mathews, T M Ahooyi, C Chen, P Mancuso, R Sariyer, P Ferrante, M Donadoni, J A. Robinson, B Sillman, Z Lin, J R. Hilaire, M Banoub, M Elango, N Gautam, R. L Mosley, L Y. Poluektova, J McMillan, A N. Bade, S Gorantla, I K. Sariyer, T H. Burdo, W Young, S Amini, J Gordon, J M. Jacobson, B Edagwa, K Khalili & H E Gendelman. Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice. Nat Comm. (2019) 10:2753, PMID: 31266936
- Su H, Cheng Y, Sravanam S, Mathews S, Gorantla S, Poluektova LY, Dash PK*, Gendelman HE*. Immune Activations and Viral Tissue Compartmentalization During Progressive HIV-1 Infection of Humanized Mice. Front Immunol. 2019 Feb 28; 10:340. (PMID:30873181) (* Shared Corresponding Author).
- Sillman B, Bade AN, Dash PK, Bhargavan B, Kocher T, Mathews S, Su H, Kanmogne GD, Poluektova LY, Gorantla S, McMillan J, Gautam N, Alnouti Y, Edagwa B, Gendelman HE. Creation of a long-acting nanoformulated dolutegravir. Nat Commun. 2018 Feb 6;9(1):443. (PMID:29402886)
- Kevadiya BD, Woldstad C, Ottemann BM, Dash P, Sajja BR, Lamberty B, Morsey B, Kocher T, Dutta R, Bade AN, Liu Y, Callen SE, Fox HS, Byrareddy SN, McMillan JM, Bronich TK, Edagwa BJ, Boska MD, Gendelman HE. Multimodal Theranostic Nanoformulations Permit Magnetic Resonance Bioimaging of Antiretroviral Drug Particle Tissue-Cell Biodistribution. Theranostics. 2018 Jan 1;8(1):256-276. (PMID:29290806)
- Gnanadhas DP, Dash PK, Sillman B, Bade AN, Lin Z, Palandri DL, Gautam N, Alnouti Y, Gelbard HA, McMillan J, Mosley RL, Edagwa B, Gendelman HE, Gorantla S. Autophagy facilitates macrophage depots of sustained-release nanoformulated antiretroviral drugs. J Clin Invest. 2017 Mar 1;127(3):857-873. (PMID:28134625)
- Zhang G*, Guo D*, Dash PK*, Araínga M, Wiederin JL, Haverland NA, Knibbe-Hollinger J, Martinez-Skinner A, Ciborowski P, Goodfellow VS, Wysocki TA, Wysocki BJ, Poluektova LY, Liu XM, McMillan JM, Gorantla S, Gelbard HA, Gendelman HE. The mixed lineage kinase-3 inhibitor URMC-099 improves therapeutic outcomes for long-acting antiretroviral therapy. Nanomedicine. 2016 Jan;12(1):109-22. (PMID:26472049) (* Shared First Author). Appeared in the cover page.
- Boska MD*, Dash PK*, Knibbe J, Epstein AA, Akhter SP, Fields N, High R, Makarov E, Bonasera S, Gelbard HA, Poluektova LY, Gendelman HE and Gorantla S. Associations Between Brain Microstructures, Metabolites and Cognitive deficits during Chronic HIV-1 Infection of Humanized Mice. Mol. Neurodegeneration. 2014 Dec 18; 9 (1): 58. (*- Equal Contribution).
- Dash PK, Gendelman HE, Roy U, Balkundi S, Alnouti Y, Mosley RL, Gelbard HA, McMillan J, Gorantla S, Poluektova L. Long-acting NanoART Elicits Potent Antiretroviral and Neuroprotective Responses in HIV-1 Infected Humanized Mice. AIDS. 2012 Nov 13; 26(17): 2135-44.
- Kadiu I, Narayanasamy P, Dash PK, Jhang W, Gendelman HE. Biochemical and Biologic characterization of Exosomes and Microvesicles as Facilitators of HIV-1 Infection in Macrophages. J Immunol. 2012 Jul 15; 189(2): 744-54.
- Dash PK, Gorantla S, Gendelman HE, Knibbe J, Casale GP, Makarov E, Epstein AA, Gelbard HA, Boska MD, Poluektova LY. Loss of Neuronal Integrity during Progressive HIV-1 infection of Humanized Mice. J Neuroscience. 2011 Mar 2; 31 (9): 3148-57.
- Eggert D, Dash PK, Gorantla S, Dou H, Schifitto G, Maggirwar SB, Dewhurst S, Poluektova L, Gelbard HA, Gendelman HE. (2010) Neuroprotective activities of CEP-1347 in models of NeuroAIDS. J Immunology. 2010 Jan 15; 184 (2): 746-56.
- Eggert D, Dash PK, Serradji N, Dong CZ, Clayette P, Heymans F, Dou H, Gorantla S, Gelbard HA, Poluektova L, Gendelman HE. (2009) Development of a platelet-activating factor antagonist for HIV-1 associated neurocognitive disorders. J Neuroimmunology. 2009 Aug 18; 213 (1-2): 47-59.
- Dash PK, Siddappa NB, Mangaiarkarasi A, Roshan PB, Aruna VM, Mahadevan A, Shankar SK, Satishchandra P, Prasad VR and Ranga U. Expanded coreceptor use of HIV-1 subtype-C molecular clones isolated from an Indian demented subject: implications for HIV-1 associated dementia. Retrovirology 2008 Mar7; 5:25.