PROJECT SUMMARY/ABSTRACT Integrative approaches, including nutrition, to address pain suffer from a paucity of mechanistic understanding, which often disproportionately impacts vulnerable populations. Affective modulation of pain in chronic disease remains an unmet need, however personalized, accessible approaches utilizing diet may provide alternative approaches for difficult-to-treat pain. Sickle cell disease (SCD) is the most commonly inherited blood disorder, primarily afflicting individuals of African and Latin American descent in the United States. Pain is a major comorbidity of SCD that can start during infancy and persist throughout life. Affective modulation in SCD is associated with social stigma, discrimination, limited resources, poor nutrition, and social isolation. Experience of pain is an outcome of somatosensory nociceptive inputs, as well as affective and cognitive state. These factors may contribute to pain by attenuating the brain’s top-down spinothalamic descending inhibitory pain pathway (DIPP) and may even interfere with response to therapy. Currently, high dose opioids are used for sickle pain. We discovered that nutrition-based approaches using nutrient- and ω-3 fatty acid-enriched “sickle diet” (SD) to target chronic pain ameliorate chronic sensory hypersensitivity and enhance spinothalamic pain inhibition. Dietary manipulation is a potent modulator of endogenous cannabinoids, such as anandamide (AEA), which has potent analgesic properties in the DIPP via dopamine (DA). DA plays a key role in the DIPP by signaling via brain regions that directly inhibit ascending pain signals in the spinal cord dorsal horn. We hypothesize that nutritionally enriched sickle diet as an early-life intervention will stimulate dopaminergic activity and ameliorate hyperalgesia in sickle mice with multigenerational benefits. In this MOSAIC K99/R00 (PAR-21- 271-010) I propose to evaluate dietary intervention as an integrative approach to address the issue of resource and nutrient limitation as factors contributing to pain in SCD. I propose the evaluation of translational endpoints as follows: K99-SA#1. Year 1-2. Use PET neuroimaging and spatial transcriptomics to evaluate the effect of SD- intervention on functional AEA and DA activity in the brain and sensory hypersensitivity of sickle mice. Milestone 1. Stratify cell type distribution in SCD DIPP with nutritional enrichment intervention. R00-SA#2. Year 3-5. Determine effects of dietary enrichment on offspring DIPP function and gene expression. Milestone 2. Establish mouse breeding colony to continue independent studies and apply techniques from K99 phase. R00-SA#3. Year 3-5 Determine teratogenicity of perinatal cannabinoid exposure in SCD offspring mice. Milestone 3. Determine associations between cannabinoid exposure and adverse outcomes and establish collaboration for epigenomic evaluation. These findings will inform the development of nutrition-based clinical approaches and will form the basis for R01 grant applications to study dietary roles in SCD pain as prospective clinical study. Deliverables. Identification of SD as a potentially effective therapy for chronic pain in SCD and evaluation of the multigenerational effects of cannabinoid exposure.

PROJECT NARRATIVE Globally, Sickle Cell Disease (SCD) is prevalent in resource-limited settings and plagued by malnutrition including for individuals living in the United States. Our goal is to identify the multigenerational effects of diet and cannabinoid exposure in the initiation and progression of pain in SCD using humanized transgenic sickle mice which show the pathobiology and features of pain similar to persons with SCD. High-throughput data and mechanisms examined herein will pave the way for clinical studies of integrative approaches to improve pain related outcomes in SCD.