Nicole M. Ashpole, Ph.D. - US grants

? DESCRIPTION (provided by applicant): The long-term career goal of Dr. Ashpole is to establish herself as a successful and well-funded, independent investigator in the field of aging, and in particular, the area of neuroscience. Thus far in her training career, Dr. Ashpole has been highly-productive with 12 peer-reviewed publications. Her doctoral training, in which she studied intracellular signaling cascades in neurons, laid a strong foundation for her current post-doctoral position. For this, Dr. Ashpole joined the laboratory of Dr. William Sonntag to study the mechanisms underlying IGF-1-dependent changes in learning and memory. Dr. Sonntag is a leading authority in the field of neuroendocrine signaling and aging. Dr. Sonntag's laboratory offers a variety of in vivo approaches which will expand her technique repertoire and allow her to become a well-rounded research scientist. The research strategy outlined in this proposal incorporates the in vitro techniques used to study signaling cascades from her doctoral training with the in vivo techniques in the Sonntag laboratory. The training program includes a mixture of laboratory training and mentoring interactions with Dr. Sonntag and an advisory council. Together, this program will ensure the Dr. Ashpole transitions to an independent investigator in the field of aging. The age-dependent loss of IGF-1 has been shown to contribute to cognitive impairment; however, the pathophysiological mechanisms underlying this effect remain to be established. The proposed studies will address this gap in knowledge by examining potential signaling pathways that may induce changes in neuronal structure and function when IGF-1 is reduced. Our preliminary data indicates that IGF-1 inhibition leads to the upregulation of the Rho-associated protein kinase (ROCK). Dr. Ashpole hypothesizes that this IGF-1- dependent upregulation of ROCK contributes to age-related impairments in learning and memory, as RhoA/ROCK activity is known to negatively influence neuronal structure and function. To better understand how IGF-1-regulated ROCK contributes to age-related cognitive decline, Dr. Ashpole will first examine the mechanisms by which IGF-1 regulates ROCK activity in Specific Aim 1. In Specific Aim 2, Dr. Ashpole will examine the effects of IGF-1-regulated ROCK on neuronal structure. Finally, in Specific Aim 3, Dr. Ashpole will examine how IGF-1-regulated ROCK influences neuronal function by measuring long-term potentiation as well as learning and memory within our mouse models. While growth hormones are often recognized solely for their contribution to cell development, it is obvious that IGF-1 plays an important role in maintaining neuronal function throughout our lifetime. Thus, this proposal addresses an area of research that is of high relevance in the field of aging. The studies that have been proposed will explore novel pathways that contribute to cognitive impairment when IGF-1 is reduced in advanced aging. Because of this, findings from this project may have long-term implications in improving the quality of life in the aging population.

Summary Approximately 50% of HIV-infected patients suffer from intractable pain and many individuals self-report the consumption of cannabis for alleviating their symptoms. However, data are still limited on the effects of cannabis in the HIV-infected, cART-treated population. The compound most studied is ?9-THC, which exerts psychoactive and addictive effects that limit therapeutic potential. Yet, there are over 120 minor cannabinoids and 200 terpenes in cannabis and our renowned Marijuana Research Laboratory has found many of them hold non-psychoactive, therapeutic effects. We identified cannabis constituents that reduce HIV-mediated inflammation/astrogliosis in cell culture and HIV-protein mediated visceral pain in mice. We hypothesize that several minor cannabinoids and terpenes will complement cART to ameliorate viremia, inflammation, and cytotoxicity caused by infectious HIV- 1. Moreover, we anticipate several of these compounds to ameliorate the inflammatory, mechanical, visceral, and neuropathic pain states promoted by cART or HIV-1 proteins. Aim 1 will determine the important cannabis constituents and targets that modulate HIV-mediated viremia and inflammation in vitro. Utilizing our extensive library of cannabinoids, terpenes, and volatile oils, we will screen a high-cannabidiol (CBD) mixture and 44 pure minor cannabinoids and terpenes against human peripheral blood mononuclear cells or microglia that are mock- infected or infected with HIV-1IIIB or HIV-1BaL. Viremia, cytokine production, and cytotoxicity will be assessed and the pharmacodynamic mechanisms will be subsequently examined using antagonists to CB1, CB2, and GPR55. Aim 2 will determine the in vivo anti-nociceptive and analgesic effects of minor cannabinoids and terpenes in rodent models of HIV-1 protein/cART-related inflammatory, thermal, visceral, and neuropathic pain. Dose- dependent anti-inflammatory, anti-hyperalgesic, and anti-nociceptive effects of cannabichromene, 10?-hydroxy- 8-tetrahydrocannabinol, and ??caryophyllene will be the pure compounds of focus, along with High-CBD extract (and other anti-inflammatory leads identified in Aim 1). These constituents will be assessed in male and female transgenic mice that express (or do not express) the HIV-1 Tat or gp120 proteins. Mice will be maintained on cART to assess potential improvements or interactions with cannabinoid-related outcomes. Aim 3 will determine the important central and enteric nervous system inflammatory mechanisms that are influenced by cannabinoids and terpenes following HIV-1 protein/cART exposure. Pain-related brain regions, spinal cord, dorsal root ganglion, ileum, edema, and plasma collected in Aim 2 will be assessed for cytokine production. Brain and ileum will be assessed for CB1 or CB2 G-protein activity via [35S]GTP?S assay as well as neuron morphology and monocyte-derived cell and astrogliosis. TThese studies will provide insight into the mechanisms by which minor cannabinoids and terpenes act, will reveal the anti-viremic, anti-inflammatory, and antinociceptive potential of non-psychoactive cannabis constituents, and will elucidate therapeutics for HIV-related and non-HIV-related, intractable pain states.