Regulation of SAMHD1 antiviral activity.

In this R01 application led by Dr. Felipe Diaz-Griffero, entitled “Regulation of SAMHD1 antiviral activity”, we are proposing to test the hypothesis that the interaction of SAMHD1 with nucleic acids is important for its anti-HIV-1 and innate immune functions. To achieve this end, we propose to utilize a CRISPR-Cas9 gene editing platform designed by my laboratory for the efficient knock-out of genes in primary human lymphocytes and myeloid cells (refer to Contribution C.1). Indeed, we have already identified conditions for the effective ablation of SAMHD1 in primary human cells (see Preliminary Data) and see no insurmountable hurdles to completing the experiments as proposed. My laboratory is located within the Fred and A. Norman Drucker Laboratory for Virology Research at Northwestern University, immediately adjacent to the labs of fellow HIV researchers Drs. Steven Wolinsky, Thomas Hope, and Richard D’Aquila. These facilities have all of the equipment and reagents required for completion of the work as outlined. Additional support from the local Third Coast Center for AIDS Research (CFAR) will ensure the experiments are carried out with maximal rigor and with input from other domain experts within the field. Dr. Diaz-Griffero and I have worked closely together in the past and currently have a paper in revision at Cell Reports. In sum, this innovative proposal has the potential to fundamentally enhance our understanding of SAMHD1 antiviral activity with broad therapeutic implications for addressing persistent virally-associated inflammation and related co-morbidities.