The renewed focus on the therapeutic potential of cannabis has sparked a significant shift in understanding the safety and efficacy of major and minor cannabinoids. Among the cannabinoids Cannabigerol (CBG) and Cannabidiol (CBD) have emerged as particularly promising compounds, each offering unique efficacy. However, the 100+ minor or rare cannabinoids are yet to be investigated comprehensively for their safety and efficacy. In this discussion, we will explore how NemaLife’s innovative platform is being used to investigate the toxicity of these promising compounds.
Rich Diversity of Cannabinoids
Cannabinoids have rapidly become one of the fastest-growing markets worldwide, yet it remains surprisingly under explored in both scientific research and regulatory frameworks. As of 2022, the global cannabinoid market is valued at an impressive $18.14 billion. However, this is just the beginning—projections indicate that the market could soar to an astounding $56.67 billion by 2030. This remarkable growth trajectory, with a compound annual growth rate (CAGR) of 15.3% over the forecast period, highlights the immense economic potential of the cannabinoid industry [2].
Cannabis sativa made its way into Western medicine in the early 19th century, marking the beginning of its recognition as a complex botanical powerhouse with significant therapeutic potential. The plant hosts an impressive array of 125 identified cannabinoids, each with distinct properties and potential health benefits. These cannabinoids are C21 terpeno-phenolic compounds unique to cannabis, making the plant a rich source of bioactive compounds with diverse effects [3].
When analyzed, these cannabinoids are categorized into several groups, reflecting the chemical diversity within Cannabis sativa. Among them, Delta-9 THC includes 25 cannabinoids, Delta-8 THC comprises 5, CBG features 16, CBD encompasses 10, CBND contains 2, CBE has 5, CBL includes 3, CBC comprises 9, CBN consists of 11, and CBT contains 9, alongside 30 other miscellaneous cannabinoids. This extensive variety highlights the plant's intricate chemical composition, far beyond the commonly studied cannabinoids like CBD and THC [3].
C. elegans as a High Throughput In vivo Model
Rodents have traditionally been the go-to model for preclinical testing of cannabinoids due to their physiological similarities to humans and well-established research protocols. However, the use of rodents in cannabinoid research comes with several significant drawbacks. One major issue is the ethical concerns surrounding animal welfare, as testing often involves invasive procedures and prolonged exposure to potentially harmful substances. Additionally, rodent studies are time-consuming and expensive, when a number of cannabinoids need to be investigated.
These limitations with mammalian models underscore the need for alternative models, such as C. elegans, which can provide more ethical, cost-effective, and translatable insights into the effects of cannabinoids. C. elegans, a tiny yet remarkable nematode, emerges as a powerful tool in overcoming the hurdles faced in cannabinoid research. Known for its rapid lifecycle, prolific reproduction, and substantial genetic similarities to humans, C. elegans offers an invaluable in vivo model for studying the physiological effects of cannabinoids. This organism's utility in research is well-documented, with contributions leading to several Nobel Prize-winning discoveries, underscoring its reliability and versatility in scientific inquiry.
What makes C. elegans particularly well-suited for cannabinoid research is its ability to model complex biological processes within a short timeframe, allowing researchers to observe the effects of cannabinoids across an entire lifespan in just a few weeks. Importantly, C. elegans possesses four major biogenic amines (octopamine, tyramine, dopamine (DA), and serotonin (5-HT)) that are responsible for modulating behavioral responses to changes in the environment, including a conserved endocannabinoid system [4].
Additionally, its transparent body facilitates real-time observation of physiological changes at the cellular level, providing detailed insights into how cannabinoids influence various biological pathways. The ease of genetic manipulation in C. elegans further enhances its value, enabling researchers to explore the specific genetic interactions involved in cannabinoid activity. As a result, C. elegans holds the potential to accelerate discoveries in cannabinoid science, offering a cost-effective, ethical, and efficient alternative to traditional models, ultimately advancing our understanding of these complex compounds.
NemaLife’s Innovative Approach
NemaLife has developed an innovative pre-clinical organism-on-chip platform for bioactives including cannabinoids. The platform combines C. elegans, microfluidics and visual AI for high throughput biology with human relevance. Microfluidics accelerates tedious nematode culture, and the visual AI auto processes millions of video images to generate phenotypic data at scale. The platform has been validated to quantify several endpoints related to toxicity as well as efficacy, all in a few weeks.
Furthermore, NemaLife’s integration of advanced imaging and AI-driven analytics ensures that even subtle effects of cannabinoids are detected and analyzed with precision. This data-driven approach enables the identification of optimal dosages and formulations that maximize health benefits while minimizing side effects. As the demand for cannabinoid-based treatments continues to rise, NemaLife’s innovative organism-on-chip platform is set to play a pivotal role in ensuring that these compounds are both safe and effective, ultimately paving the way for their broader acceptance and use in clinical practice.
Application of NemaLife’s Organism-on-Chip Platform for Cannabinoid Research
In collaboration with our partners, NemaLife’s organism-on-chip platform has been used to study the long-term safety of cannabinoids. Below we describe some of these collaborative studies, which have shown the significant potential of our platform for adding pre-clinical evidence to cannabinoid safety and efficacy.
One of the most well-known cannabinoids, CBD, has garnered significant attention in scientific research, second only to THC in its prominence. Despite the numerous studies conducted on CBD, there had been a notable absence of lifelong toxicity studies until recently. Using the NemaLife platform, both acute and long-term exposure studies of CBD to adult C. elegans at physiologically relevant concentrations were conducted [5]. No lethality was observed when exposed to 0.4-4000 μM CBD in the acute exposure paradigm. Notably, whole-life exposure of C. elegans to 10-100 μM CBD revealed a maximum life extension of 18% observed at 40 μM CBD. These results show the importance of a short-lived in vivo model to generate lifelong exposure data.
Likewise, CBG, often regarded as a minor cannabinoid due to its typically low concentration in most cannabis strains, has recently garnered increased attention for its therapeutic potential and favorable safety profile. Using the NemaLife platform, both acute and long-term exposure studies of CBG to adult C. elegans at physiologically relevant concentrations were conducted [5]. No lethality was observed when exposed to 0.075 μM - 3.75 mM CBG in the acute exposure paradigm consistent with a 14-day toxicity study in rodents. Notably, lifelong CBG administration resulted in mean lifespan extension of 13.9%–19.1% in C. elegans.
Figure 4: Tim Lefever Ph.D., the Director of Discovery at Nalu Bio presented findings from NemaLife’s organism-on-chip platform during a talk titled “Characterization of Less Studied Cannabis Phytochemicals” at the CannMed 2024 Conference. You can also watch the full video of the presentation here: https://lnkd.in/gXGtUnYZ
In a recent collaboration effort with Nalu Bio, NemaLife’s organism-on-chip platform was used to evaluate chronic toxicity and neuroactivity of minor or rare cannabinoids. A total of 27,000 worm-subjects and more than 2 million images were analyzed to generate in vivo phenotypic data. More research is being conducted with this dataset to illuminate the safety and efficacy of minor cannabinoids.
Conclusion
The ongoing research into cannabinoids is gradually peeling back the layers of mystery that have long surrounded these compounds. With their complex chemistry, lack of significant toxicity in early studies, and potential health benefits, cannabinoids are poised to play an increasingly important role to address human health conditions.
Through its innovative approaches, NemaLife can support the safe integration of cannabinoids into treatments, ensuring that these compounds are utilized to their fullest potential in enhancing human health and well-being. This comprehensive approach positions NemaLife at the forefront of cannabinoid research, poised to unlock new opportunities and drive the future of bioactive discovery and development in this rapidly evolving field.
References:
1. Gülck, T., & Møller, B. L. (2020). Phytocannabinoids: Origins and Biosynthesis. Trends in Plant Science, 25(10), 985-1004.
2. U.S. cannabinoids market size, Share & Trends report, 2030. U.S. Cannabinoids Market Size, Share & Trends Report, 2030. (n.d.). Source
3. Radwan MM, Chandra S, Gul S, ElSohly MA. Cannabinoids, Phenolics, Terpenes and Alkaloids of Cannabis. Molecules. 2021 May 8;26(9):2774. doi: 10.3390/molecules26092774. PMID: 34066753; PMCID: PMC8125862.
4. Estrada-Valencia R, de Lima ME, Colonnello A, Rangel-López E, Saraiva NR, de Ávila DS, Aschner M, Santamaría A. The Endocannabinoid System in Caenorhabditis elegans. Rev Physiol Biochem Pharmacol. 2023;184:1-31. doi: 10.1007/112_2021_64. PMID: 34401955; PMCID: PMC8850531.
5. Land MH, Toth ML, MacNair L, Vanapalli SA, Lefever TW, Peters EN, Bonn-Miller MO. Effect of Cannabidiol on the Long-Term Toxicity and Lifespan in the Preclinical Model Caenorhabditis elegans. Cannabis Cannabinoid Res. 2021 Dec;6(6):522-527. doi: 10.1089/can.2020.0103. Epub 2020 Nov 20. PMID: 33998871; PMCID: PMC8713279.
6. Kulpa J, Lefever TW, Trexler KR, Henderson RG, MacNair L, Toth ML, Vanapalli SA, Rahman M, Gupta S, Bonn-Miller MO (2023) Toxicity of cannabigerol: examination of long-term toxicity and lifespan in Caenorhabditis elegans and 14-day study in Sprague Dawley rats, Cannabis and Cannabinoid Research 8:S1, 62–70, DOI: 10.1089/can.2023.0035.
Comments