Session : The Cannabis Plant: A Sustainable Source of Bioactive Molecules and New Pre- and Post-Biotics of Nutritional and Medical Importance

Omega-3 fatty acids support cardiometabolic health and reduce chronic low-grade inflammation. These fatty acids may impart their health benefits partly by modulating the endocannabinoidome and the gut microbiome, both of which are key regulators of metabolism and the inflammatory response. Whole hemp seeds (Cannabis sativa) are of exceptional nutritional value, being rich in omega-3 fatty acids, especially stearidonic acid (SDA).  Supplementation of a high-fat, high-sucrose diet with whole hempseeds did not affect weigh gain, adiposity, or food intake, whereas linseed substitution did. Furthermore, hemp seed substitution mitigated diet-induced obesity-associated increases in intestinal permeability and circulating PAI-1 levels, while having no effects on markers of inflammation in epididymal adipose tissue, which were, however, increased in mice fed linseeds. Both hemp seeds and linseeds were able to modify the expression of several endocannabinoidome genes and markedly increased the levels of several omega-3 fatty acid–derived endocannabinoidome bioactive lipids with previously suggested anti-inflammatory actions in a tissue specific manner, despite the relatively low level of seed substitution.  Interestingly, incubation of both preadipocytes and adipocytes with SDA resulted in the endogenous production of the novel endocannabinoidome lipids 2-stearidonoyl glycerol and N-stearidonoylethanolamine (SDEA), which, while not modulating lipid levels in adipocytes, did decrease lipid levels in hepatocytes, suggesting that these compounds have lipolytic activity, in line with their ability to activate peroxisome proliferator activator alpha (PPARA)-mediated transcription.  Furthermore,  utilizing an in vitro culture system of the human gut microbiome, we show that SDA-rich oil modulates the bacterial populations in a host-specific manner resulting in microbially-produced SDEA.  Taken together, these results suggest that hemp seed-containing foods might represent a source of healthy fats that are not likely to exacerbate the metabolic consequences of obesogenic diets while producing intestinal permeability protective effects and some anti-inflammatory actions, potentially through the production of SDA-derived endocannabinoidome bioactive lipids by host tissues and/or gut microbiota.

The Western diet, consisting of highly palatable and caloric food (HPF), represents the main environmental cue triggering addictive-like behaviors and obesity by inducing a long-lasting alteration in synaptic plasticity in mesolimbic dopaminergic circuits associated with reward responses. Studies, in both rodents and humans, have suggested that the gut microbiome is modulated by the diet and intervenes in the control of consumption of, and dependence from, HPF. These effects are partly modulated by endocannabinoidome (eCBome) signalling, in both the brain and gut. Studies about the effects of phytocannabinoids and endocannabinoid-like molecules on the morpho-functional integrity of the gut barrier and on central neurogenesis, as well as the role of oleic acid-derived mediators N-oleil-glycine and N-oleil-alanine on the gut microbiota composition in mice will be presented, in order to discuss about new nutritional strategies aimed at reducing the rewarding and obesogenic properties of HPF.

The food products derived from Hemp (Cannabis sativa subsp. sativa) are steering a large sector market that will reach almost USD 5 billion by 2026. Hemp seeds are considered a nutritional powerhouse, source of fibers and unsaturated lipids. During the food processing chain, the external part of the seed is discarded, although it contains a high number of proteins. This material should be transformed into value-added products to meet the demand of sustainable source of proteins and to reduce food waste. Hemp seed bran has shown to have prebiotic effects and to be rich in bioactive compounds, suggesting having potentials as a food product or fiber supplement. Notwithstanding, information on the functional capabilities of hemp seed bran is still little and the study of its impact on human colon microbiota would pave the way to comprehension. In fact, the human colon microbiota is the main responsible for digestion of vegetable fibers and the only for digestion of prebiotics. Recently with the aid of omics the precise characterization of prebiotic potential of hemp seed bran and its hydrolyzed product was revealed. The hemp seed bran samples were compared to a prebiotic positive control, the fructooligosaccharides (FOS), after human colonic fermentation using an in vitro colon gut model. During fermentation, metabolomics (SPME GC/MS) and microbiomics (MiSeq and qPCR) analyses were conducted. The results highlighted the functional properties of hemp seed bran towards human colon microbiota by the observation of some ecological indicators. For example, hemp seed bran produces beneficial organic fatty acids, as the medium chained ones that are considered an indicator of colon eubiosis, reduces more detrimental phenol derivates, as p-Cresol, that is considered an indicator of negative proteolytic fermentation, and produces bioactives molecules as 4-Terpineol, that is a potent antioxidant terpene. Considering microbiomics, hemp seed bran fosters the growth of beneficial bacteria as Bifidobacterium bifidum; and limits opportunistic bacteria as Bilophila wadsworthia, that usually induce dysbiosis by sulfate reduction and H2S formation. The results collected evidenced the functionality and safety of hemp seed bran towards the human colon microbiota with an in vitro gut model approach. So far, following the principles of OneHealth and 3Rs it is possible to valorizes a byproduct as a sustainable protein source and contribute to the reduction on animal testing in food science.

Cannabis is widely used for medicinal and recreational purposes, however, there is a lack of understanding about its effects on human physiology, medical outcomes, and concerns about consistency and reproducibility of the derived products. These concerns are driven primarily by the inherent chemical complexity of a biological organism. What makes whole-plant extracts more challenging is that they are not just chemically complex, but also chemically variable. This variation is due to a combination of the genetic background of the plant, as well as the environment in which it is grown. In order to produce reproducibly consistent extracts, it is critical that they be derived from genetically stable plants grown under uniform conditions. Then, and only then, will it be possible to conduct pre-clinical and clinical evaluation to fully understand these complex interactions and biological outcomes. Here, I will talk about our pipeline to develop uniform and genetically stable cannabis plants.

Introduction: Inflammatory bowel diseases (IBDs) are associated with alterations of various members of the endocannabinoidome (eCBome) and gut microbiome, both of which hold therapeutic potential for the treatment of inflammation and affective disorders associated with IBDs, such as ulcerative colitis. The endocannabinoidome and the gut microbiome are strongly influenced by diet and thus may be targeted with nutritional supplements.

Methods: We assessed the effects of: 1) a nutritional supplement formulation (Form) comprised of a proprietary blend of cannabimimetic plant extracts (produced by Silicycle Inc.), or 2) omega-3 fatty acid-rich fish oil (FO) or seal oil (SO), on the development of inflammation, intestinal permeability, anxiety, the gut microbiome and colon eCBome in a dextran sodium sulfate (DSS)-induced mouse model of colitis. Mice were pretreated with supplements for 2 weeks prior to induction of a 5-day DSS-treatment protocol; behavioural assessments were performed 7 and 8 days after initiation of DSS and mice were sacrificed, and samples harvested on day 9.

Results and Conclusions: Targeting the eCBome with a proprietary blend of cannabimimetic plant extracts or omega 3 fatty acid-rich seal oil resulted in significant improvement in intestinal permeability in DSS-treated mice, with the formulation also improving some select signs of anxiety-like behaviour. DSS-induced colitis altered the colonic bacterial community, and while none of the treatments significantly affected this, changes in specific taxa were observed. All treatments, but particularly the administration of oils, resulted in the inhibition of the DSS-induced increase in the abundance of the family Eryspelatoclostridiaceae and decrease in the abundance of the genus Family XIII UCG001, alterations of which have been associated with colitis. DSS-induced decreases in AEA and increases in 2-AG levels within the colons of mice were inhibited by the cannabimimetic formulation but not omega-3 fatty acid-rich oils, suggesting that modification of classic endocannabinoid levels may be a mechanism by which the formulation improved physiological and behavioural parameters indicated above. Conversely, FO and SO increased the levels of EPA-derived PGE3, suggesting that FO- and SO-mediated improvements in gut barrier permeability may be linked to this anti-inflammatory prostaglandin. Effects of these treatments on colonic inflammation and plasma and brain eCBome levels and inflammation markers are ongoing.

Despite the long history of cannabis cultivation, industry faces many unique regulatory and practical challenges in growing and using this crop.  The wide variety of cultivars developed, many most recently for the recreational market, but also those producing non-euphoric cannabinoids, has resulted in growers, and those interested in developing alimentary, nutraceutical and/or pharmaceutical cannabis products, having to make many choices as which cultivars to use.  Not the least of which, is taking into account how the variety of bioactive phytocannabinoids produced by a plant interacts with the human body, for example by affecting our own “endogenous cannabis signalling system”, the endocannabinoid system.  This panel of industry-leading experts will provide insights into the realities of working with cannabis.