Pediatric Latent HIV Reservoir Characterization and Quantification Assay

PROJECT SUMMARY ABSTRACT Public Health Problem Modern antiretroviral therapy (ART) combinations greatly improve survival of HIV infected individuals and reduce HIV transmission. However, there are still 38 million people living with HIV (PWH) worldwide and of those 1.8 million are children under the age of 15, highlighting the imperative need for curative therapies. In limited

resource areas, where pediatric HIV is most common, the use of new assays that require less blood volumes, lower cost and less labor-intensive methods are necessary to facilitate clinical trials aimed at reducing the size of the latent reservoir and/or to achieve a cure or ART-free remission. The ultimate value of the SBIR Phase I project will demonstrate the ability of our technology to characterize and

quantify the pediatric HIV-1 reservoir, which remains a critical unmet need in the HIV cure field. Issues with Current Solutions & How Product Meets Unmet Needs RNAamp allows direct detection of RNA, which provides the most accurate measure of the HIV replication competent reservoir. The assay is proven to be faster and have better reliability and quantification than HIV

reservoir assays based on QVOA, RT-qPCR, RT-ddPCR, iSCA and IPDA. RNA target amplification and detection is accomplished by oligonucleotide-templated photoreduction using short, sequence-specific profluorophore and catalytic probes. Reproducibility is outstanding (4% variance between replicates) and Limit

of Detection (LoD) is 1 HIV-infected cells/1 x 106 PBMCs or 1-10 RNA copies/mL, using a standard blood draw (10-30 mL). Jan Biotech’s RNAamp quick, direct RNA test, with its use of small blood samples, ease of sample preparation, reliability and sensitivity, quantitative ability and short time to result, is ideally suited for characterization and

quantification of the rebound-competent pediatric HIV reservoir. Summary of Approach Jan Biotech has developed an assay with a prediction accuracy of 0.92 for time to rebound after treatment interruption for early/acute-treated adult participants. The assay, RNAamp, utilizes nonezymatic, chemical amplification of cell-associated HIV RNA targets to provide quick, sensitive, and quantitative measurement of

the HIV reservoir. No other assay, including QVOA, IPDA, iSCA, RT-qPCR, or RT-ddPCR, provided a prediction accuracy of greater than 0.2 for the same adult participants. This proposal is to develop RNAamp for cross- subtype (i.e., non-B subtype) HIV reservoir detection, specifically for pediatric testing, for which the use of small

samples easily tested by RNAamp is crucial. Pediatric HIV-infected individuals are typically treated during early HIV infection, often treated at birth, and physician-monitored or spontaneous treatment interruption is more frequent in this population. Pediatric treatment efficacy and treatment interruption clinical trials typically include

non-B subtype HIV-infected participants. Collaborators and Unique Resources Jan Biotech, Inc., with expertise in molecular diagnostic development, will consult with Jonathan Li, MD, MMSc Assistant Professor of Medicine, Harvard Medical School, in a AIDS Clinical Trials Group (ACTG) study and Harris Gelbard, MD, PhD, University of Rochester Medical School.

Phase I Specific Aims Specific Aim 1: Develop HIV-1 cross-subtype (CS) RNAamp multiplexed assay Specific Aim 2: Correlate CS-RNAamp to Pol RT-qPCR using non-B subtype HIV-infected cells Specific Aim 3: Pediatric human sample testing using CS-RNAamp HIV reservoir characterization Market after Phase II Completion

Both Gilead and Merck are commercially interested in the technology. Regulatory approval will be sought for commercialization.