Background-Significance-Hypotheses: Cannabis is consumed for medicinal and recreational purposes; however, impact of consumption on diseases associated with compromised gut barrier function and the underlying mechanisms involved processes remain unclear. We will test the hypothesis that cannabis provides protection against gut barrier dysfunction in metabolic and inflammatory bowel diseases. We aim to identify the impact and possible differential effects of cannabis exposure on gut barrier function in two robust murine disease models that display moderate vs. severe gut barrier dysfunction (i.e., diet-induced obesity and experimental colitis).

Innovation: To achieve the goals of our study, several key innovative approaches and novel methodologies are employed including use of two distinct mouse models associated with moderate or severe gut barrier dysfunction, optimization of UPLC-MS/MS methods for quantitating levels of delta-9 THC in cannabis and conversion of precursors to delta-9 THC via optimized chemical reactions that mimic consumption of cannabis in humans (i.e., heating of material), and (iii) optimization and utilization of reliable and reproducible assays of gut barrier function.

Design-Methods: We will administer cannabis extracts (5 mg per kg delta-9 THC content) (n=12 each) of male and female IntCB₁R-/-mice and IntCB₁R+/+ control mice (i) maintained on a low-fat/sugar or high-fat/sugar diet for 60 days or (ii) subjected to DSS-induced colitis.

Study Design: 2 drug conditions (whole cannabis extracts + vehicle) x 2 genotype (IntCB₁R-/-, controls) x 2 diets (standard, western diet; Aim 1) or colitis (DSS, controls with no DSS; Aim 2) x 2 sex x 12 mice per condition = 384 total mice.

Statistical Plan of Analysis: Power analyses for determining required sample size for each group is based on preliminary data. We will analyze all associated data by separate 2-way ANOVA with Tukey multiple comparisons tests for individual drug treatments (cannabis extract vs. vehicle) vs. genotype (IntCB₁R-/-, controls). Separate analyses will be made for each individual sex as well for each individual diet or colitis condition (DSS or control). Analyses will be made for effects on markers of gut barrier function and inflammation.

Implications: These studies will support evidence-based public policy associated with cannabis use and identify possible therapeutic benefits for treating disorders that affect the gastrointestinal tract.