¹Division of Clinical Pharmacy, School of Pharmacy, The Hebrew University, Israel

²Department of Psychiatry, Hadassah Hebrew University Medical Center, Israel

³Mental Health Division, Ministry of Health, Israel

⁴Department of Anesthesia, Hadassah Hebrew University Medical Center, Israel

⁵Israeli Ministry of Health Department for the Treatment of Substance Abuse, Ministry of Health, Israel
 

 

 

Background: Although most cannabis use is recreational, cannabis is already approved for various medical indications. One of most controversial safety concerns regarding the use of cannabis is its association with mental illness, in particular, induction or exacerbation of psychosis and schizophrenia, cognitive compromise in adolescents, suicidal attempts and depression. That said, current available data concerning mental illness outcomes with regards to cannabis exposure is based mostly upon cross-sectional studies of recreational users of cannabis and is therefore subject to inherent limitations and weaknesses. Therefore, benefits and risks of medical cannabis use should be evaluated by meticulously designed studies comparing large and well-matched populations. The proposed study will enable, for the first time, to explore favorable and deleterious effects of medical cannabis drawing on a large population registry. Findings from the study will provide crucially needed data that could be immediately implemented in clinical practice, will contribute to the ongoing debate about legalization of recreational cannabis and may lay the groundwork for additional research.

Study objectives: To study the association between exposure to medical cannabis and the risk for mental illness, including the risk for psychotic episodes, anxiety and suicidal behavior in previously mentally healthy subjects, as well as prompt a relapse of these conditions in individuals with psychiatric history.

Data acquisition and Study design:  

Data will be retrieved from three databases. The first is the Israeli Medical Cannabis Agency at the Ministry of Health. This database contains the names of all patients who were granted a license to use medical cannabis, the indications and symptoms for cannabis usage and the monthly and total amount dispensed, Socioeconomic grade based on AGAS. This database will be linked with databases extracted from computerized records of Health Service Providers containing socio-demographic information, medical diagnoses, medication, and additional clinical data, and the Israel Ministry of health Psychiatric hospitalizations database, documenting psychiatric hospitalizations and key measurements related to these hospitalizations.

Subjects exposed to medical cannabis will be compared to a clinically matched unexposed control group (excluding Post Traumatic Stress Disorder [PTSD] patients) in a cohort study design. We will compare the incidence rate and hazard ratios in the exposed group to the unexposed control group, adjusting for potential confounders.

We will also perform subgroup analysis: a subgroup of patients treated with medical cannabis for the indication of PTSD and pain will be matched to a control group consisting of patients with PTSD treated with opioids for pain, by high dimensional propensity score.

Another subgroup analysis will be performed on medical cannabis exposed patients in a case-crossover design. The combined information will be analyzed after encryption for data privacy purposes to conclusively determine the relationship between exposure to medical cannabis and mental illness.

Importance to Medicine: Findings from the study will provide crucially needed data that could be immediately implemented in clinical practice, will contribute to the ongoing debate about legalization of recreational cannabis and may lay the groundwork for additional research.

 

 

 

 

¹Division of Clinical Pharmacy, School of Pharmacy, The Hebrew University

²Department of Psychiatry, Hadassah Hebrew University Medical Center

³Hebrew University-Hadassah Braun School of Public Health and Community Medicine, Jerusalem   

 

 

⁴Department of Anesthesia, Hadassah Hebrew University Medical Center

⁵MoreBetter LTD

 

 

 

Although cannabis products are marketed for mental-related conditions, one of the main concerns of cannabis use is its adverse mental effects. Studies suggest an association between cannabis use and the risk for mental diseases or exacerbation of existing mental conditions. This database study characterizes users who experienced negative mental emotions after cannabis exposure.

Methods: We used a self-report database of the ReleafApp by MoreBetter LTD, which enables users to track real-time cannabis user experience. From a list of 42 possible emotions and symptoms  on which the user may report, we focused on a group of negative mental emotions as an outcome that may indicate mental deterioration. Multivariable logistic regression models with mixed effects analyses were performed to study the association between user’s traits, cannabis products, and reporting negative mental emotions.

Results: 144,954 sessions of cannabis consumption were reported by 6,191 users. Females and non-binary gender were associated with an increased risk for negative mental emotions than males (OR=1.16, 95%CI: 1.05-1.28, OR=1.48, 95%CI: 1.26-1.34). Older age was associated with less negative mental emotions than young age (OR=0.88, 95%CI: 0.78-0.99, OR=0.71, 95%CI: 0.62-0.81, in 30-40 and 40-50, respectively compared to the age group of 18-30 years). New users were associated with an increased risk for negative mental emotions compared to experts (OR=1.35, 95%CI: [1.12-1.64]). Oral cannabis products were associated with negative mental emotions compared to flowers. Nevertheless, the experience of negative mental emotions was not associated with a change in the response to the cause for which the cannabis was used. In a cluster analysis, negative mental emotions were more correlated with one other than with other emotions. 

Conclusions: Cannabis use in females and non-binary users was associated with negative mental emotions. Oral cannabis products are also associated with negative mental emotions, compared to flowers products. Older age and previous experience with cannabis were associated with fewer negative mental emotions. Further studies should examine the abovementioned traits in the context of cannabis use and mental illness.

 

 

 

Division of Clinical Pharmacy, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel.

Department of Development and Regeneration, KU Leuven, Leuven, Belgium 

Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Leuven, Belgium 

Department of Hospital Pharmacy, Erasmus MC University Medical Center, Rotterdam, the Netherlands 

Clinical Pharmacology Unit, Shamir Medical Center (Assaf Harofeh), Zerifin, Sackler Faculty of Medicine, Tel-Aviv University, Tel Aviv, Israel

The Autism Center, Alut, Shamir Medical Center (Assaf Harofeh), Zerifin

 

 

Despite the increased use of medical cannabinoids, the efficacy and safety of the treatment among children remain uncertain. The objective was to study the efficacy and safety of medical cannabinoids in children. The search included studies through 11-May-2020. Selection criteria included studies evaluating efficacy and safety outcomes of medical cannabinoids (tetrahydrocannabinol, cannabidiol and other cannabis derivatives) versus control in children, independently assessed by two reviewers. Eight studies were included, all of which are randomized controlled trials. Cannabidiol is associated with 50% reduction in seizures rate (Relative Risk (RR) = 1.69, 95% CI [1.20–2.36]) and caregiver global impression of change (Median Estimated difference = (− 1), 95%CI [− 1.39–(− 0.60)]) in Dravet syndrome, compared to placebo. While cannabidiol was associated with a reduction in reported seizure events (RR = 0.59, 95% CI [0.36–0.97]), no association was found in products contained also tetrahydrocannabinol (RR = 1.35, 95% CI [0.46–4.03]). Higher dose of cannabidiol was associated with decreased appetite (RR = 2.40, 95% CI [1.39–4.15]). A qualitative assessment suggests that medical cannabinoids might be associated with adverse mental events. In conclusion, cannabidiol is associated with clinical improvement in Dravet syndrome. However, cannabidiol is also associated with decreased appetite. Adverse mental events were reported as well, however, more research should be performed to assess well this outcome.

 

 

In recent years, the use of Cannabis in the adult population has dramatically increased due to legalization of Cannabis in parts of the United States and the use of Cannabis or Cannabis extracts as palliative medication. One of the main barriers hindering the use of Cannabis flowers as medicinal agents is the lack of uniformity in terms of the content of active ingredients. Moreover, Cannabis in nature is highly pollinated, which is the reason for extremely inhomogeneous plant material, hence obtaining new hybrid seeds with consistent genetic makeup is almost an unreachable goal. Furthermore, to stabilize the active ingredients’ concentration in Cannabis plants is even more challenging since it is affected by various factors: plant genetics, growing and storage conditions, the state of maturity at harvest, etc. This is the major reason for avoiding producing Cannabis seeds worldwide and using vegetative propagated plants instead. All these reasons together cause difficulty in the repeatability of the patient’s dosage. Thus, in order to promote Cannabis flowers as validated medicine, the ability to quantify the exact percentage of active ingredients in the plant is required.

The active ingredients of cannabis plants are the cannabinoids, a class of diverse chemical compounds that are concentrated in specialized glandular structures called trichomes. The primary cannabinoid is phytocannabinoid tetrahydrocannabinol (THC) while cannabidiol (CBD), another cannabinoid, is the primary analgesic compound of Cannabis. In addition, there are at least 85 additional plant cannabinoids having varied physiological effects. [1]. Gas Chromatography (GC) and High Performance Liquid Chromatography (HPLC) are commonly used approaches for quantification of plant cannabinoids.[2] Although those approaches can be used to type Cannabis strains according to THC and CBD quantities, they require processed plants’ material and are time consuming. Moreover, they can only be implemented on mature flowers, which requires growing a diversity of all plants before selection can be made. Therefore, in order to promote cannabis as medicine there is a need to act on two parallel platforms; stabilization of the active ingredients in the cannabis plant while producing the ability to quantify the percentage of active ingredients in each flower. One possible approach for quantifying active ingredients in whole (unprocessed) plants or production of Cannabis seeds and breeding acceleration process might be achieved by NIR technology.[3] NIR spectrometry supported by image analysis and machine learning can be used to detect and quantify cannabinoids in whole plants or plant material by remote sensing[4]. Recently, my laboratory has revealed the ability to accurately analyse THC levels in Cannabis flowers by NIR spectroscopy (Fig.1). NIR spectrometers/cameras record the absorbance/reflectance spectrum of samples irradiated with light at wavelengths between 700 nm and 2500 nm. NIR radiation is highly penetrative and can be applied to a sample without any preparation/destruction. Although the resulting absorbance or reflectance spectra is not highly discriminative, it can be used to quantify active ingredients and other agricultural features such as germination rate, male/female plants, disease resistance etc., by using NIR calibration models. This new system technology based on NIR spectroscopy will be able to determine cannabinoids content in Cannabis flowers as well as genetic features in Cannabis seeds, and most notably the active ingredients ratio. By this novel technology, we will be one-step ahead towards implementation of consistent cannabis flowers for medical use. Moreover, we will achieve a shorter and much cheaper breeding process, which can cause a dramatic change in the entire plant breeding industry.
 

REFERENCES

1. El-Alfy, Abir T.; Ivey, Kelly; Robinson, Keisha; Ahmed, Safwat; Radwan, Mohamed; Slade, Desmond; Khan, Ikhlas; Elsohly, Mahmoud; Ross, Samir (2010). "Antidepressant-like effect of Δ9-tetrahydrocannabinol and other cannabinoids isolated from Cannabis sativa L". Pharmacology Biochemistry and Behavior 95 (4): 434–42.
2. F.E. Dussy, C. Hamberg, M. Luginbuhl, T. Schwerzmann, T.A. Briellmann, Forensic Sci. Int. 149 (2005). Isolation of Δ9 -THCA-A from hemp and analytical aspects concerning the determination of Δ9 -THCin cannabis products. Forensic Science International 149(1):3-5.
3. T. H. Reijmers; C. Maliepaard; H. C. Van Den Broeck; R. W. Kessler; M.A.J. Toonen; H. Van Der Voet (2005). Integrated statistical analysis of cDNA microarray and NIR spectroscopic data applied to a hemp. Journal of Bioinformatics and Computational Biology. 3(4): 891-913.
4. I. Azaria.; N. Goldshleger.; E. Ben-Dor.; R. Bar-Hamburger (2009). Detection of Cannabis Plants by Hyper-Spectral Remote Sensing Means. Tel Aviv University Publication.

Waste management is a huge concern for the Cannabis growers' community. After the female Cannabis flowers are harvested the growers are left to dispose of all the vegetative parts. Legal growers must document every stage of plant management that includes plant material that is being discarded. Thus, Cannabis disposal is subjected to strict waste regulations. Furthermore, traditionally the majority of crop residues are used as animal feed but the strict regulations on Cannabis residues are preventing this use.

 

One way to manage this crop waste is the incorporation of crop amendments into the soil (green manure). Ploughing plant debris into the soil to generate green manures has been proposed to affect plant health and to control diversity of soil borne and foliar pathogens. My lab expertise in plant defence mechanisms  together with our ability to use identified characteristics (e.g. fluorescein diacetate, substrate respiration and biocontrol agent population)  that correlates with green manure diseases suppression will advance  us towards the calibration of Cannabis waste as green manure.

In our laboratory we also study the accumulation of plant secondary metabolites during plant defence responses. Since it is known that developmental stages and environmental conditions affect the metabolite profiling in Cannabis, we plan to study the suppressive effect of cannabis metabolites throughout the different developmental stages and under different conditions on Botrytis cinerea and other plant pathogens. We will later use those findings to calibrate cannabis waste as suppressive green manure for the control of soil borne pathogens and for metabolic engineering in order to confer resistance against B. cinerea.

 

Keywords: green manure, plant pathogens, metabolic engineering, secondary metabolites.

Department of Plant Pathology and Microbiology, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel

 

Hybrid varieties of Cannabis Sativa, Cannabis Ruderalis and Cannabis Indica are used as drug crops. These crops produce large and dense flowers which are very attractive to Botrytis cinerea - the gray mold disease causal agent. B. cinerea is the most significant pathogen on Cannabis plants and its infection results in enormous economic losses due to yield damages. When Cannabis is grown in-doors under high humidity and in high density of large moisture-retaining female buds, B. cinerea can reach epidemic magnitudes and completely destroy a Cannabis crop within a week. Drug varieties are most susceptible during flowering near harvest time as growers cannot use any chemical fungicides due to consumers' health concerns. Thus, developing new strategies to control this pathogen is essential if we wish to secure Cannabis crops for the health of both consumers and the environment.

In my laboratory, we are focusing on various aspects of the plant's defence responses against the fungal plant-pathogen Botrytis cinerea. To cope with pathogen attack, plants have evolved a wide range of physical, molecular and biochemical defence mechanisms. We target to refine the multi-layer defence responses against B. cinerea by studying the molecular and biochemical interactions of B. cinerea with the plant's defence response.

Our research in the laboratory revealed diverse knowledge on B. cinerea – host interaction and plant induced resistance mechanisms against this pathogen. We also isolated a unique and efficient biocontrol agent that uses a complex mode of actions against B. cinerea including antibiosis and activation of the plant defence response. We aim to use these findings to enhance the defence response of Cannabis plants in order to control B. cinerea and other pathogens infecting this crop. 
 

 

Keywords:Botrytis cinerea, plant immunity, pathogens, fungi, resistance. 

Genome editing using the CRISPER/Cas9 technology opened new possibilities in research and in biotechnology for many organisms including plants. To date, targeted gene knockout using the CRISPER/Cas9 system has been used in many plant species, with high efficiency. Despite the high potential of the system to manipulate genes and to serve as a tool in breeding programs, there is still no published work on using CRISPER/Cas9 in cannabis. We believe that genome editing can be a powerful tool in generating new cannabis strains having desirable characteristics. To establish the genome editing system in cannabis, we are developing a method for transient transformation in protoplast with a plasmid carrying the Cas9 gene and the guiding molecule. This method allows the Cas9 to induce a mutation without integrating to the genome, generating a non-transgenic cell, with the desirable mutation.  Following the transformation, the protoplast are induced to regenerate shoots, resulting in a mutated plant that is unify (non-chimeric) and true-to-type. Calibrating this method to exhibit high efficiency will enable us and other to generate many new plants in short time in a non-transgenic manner. There are many traits that can be manipulated by gene editing. For example, creating plants with agro-technical advantages, or manipulating the cannabinoid biosynthesis pathway.  

• Key words: Genome editing, Plant regeneration, Transient transformation

Evidence for the use of Cannabis Sativa as a treatment for pain can be traced back to the beginnings of recorded history. To date, the main use of ‘medical marijuana’ is for treating the most debilitating chronic pain disorders, such as neuropathic and cancer pains. However, the molecular and cellular mechanisms by which cannabinoids reduce pain are not known. While two cannabinoid-specific receptors (CB1 and CB2) have been identified, their pharmacological or genetic blocking did not abolish the effect of cannabinoids. This points to other, yet unidentified, receptors that mediate cannabinoid-induced analgesia. Recently, a family of somatosensory TRP channels, specifically TRPV1 and TRPA1, was proposed to be the cannabinoids ionotropic receptors. These receptors are mainly expressed on nociceptive fibers (i.e. Ad- and C- fibers) and their short activation evokes nociceptive pain, while their prolonged activation results in neuronal ‘shut-down’ and analgesia. In this multidisciplinary collaborative project, we are using specific pharmacological tools together with ion imaging, electrophysiology and behavioural tests to establish the role of TRPV1 and TRPA1 channels in cannabinoids-induced analgesia. We believe that detailed comprehension of mechanisms of cannabinoids-induced analgesia could facilitate rational design of novel and specific analgesics.

This study will examine the effect of Cannabis administered to patients who are admitted to the emergency room suffering from acute radicular pain. In a blind, randomized & controlled study patients will receive either Cannabis or a placebo. They will then be monitored for 24 hours for the observation of multiple parameters.

Cannabis is consumed recreationally or for medical reasons in 5–15% of the Israeli population; however, the perioperative effects of cannabis use on anesthesia and surgery are poorly understood.

We have received a grant and preliminary IRB approval to study the effects of a cannabis extract as anesthetic premedication on perioperative anxiety, and on postoperative pain, nausea, and vomiting.

We are planning a national study with a patient population of approximately
16,000 individuals that has received Ministry of Health approval for the use of medical cannabis in Israel, in collaboration with Prof O. Bone of Hadassah’s Department of Psychiatric Medicine and Prof Y. Neumark, Director of the School of Public Health. The question we are trying to address is: Does cannabis consumption increase mental sickness?  

Traumatic brain injury (TBI) is the leading cause of death in the young age group and the most commonly identified cause of epilepsy in adult populations older than 35 years. It triggers a cascade of events characterized by the activation of molecular and cellular responses, mostly harmful, leading to secondary injury. Parallel to these deleterious processes, neuroprotective events also take place including secretion of growth factors, and activation of anti-apoptotic signalling pathways. Despite the acute need of an effective pharmacological means to treat TBI victims, pharmaco-therapy remains scarce and new drug targets are clinically necessary. Previously it was shown that treatment with synthetic cannabinoid 2-AG, attenuated edema formation, infarct volume, blood-brain barrier permeability, neuronal cell loss at the CA3 hippocampal region and neuro-inflammation following closed head injury (CHI). Moreover, improved recovery of neurobehavioral function was noted for up to 3 months after treatment of CHI mice with 2-AG. Based on these studies it is clear that the cannabinoid system plays a critical role in neuroprotection following TBI. However, little is known about the role of the cannabis plant in TBI. Our preliminary results indicate that mice treated with one of the major compounds that were found in the cannabis plant, cannabidiol (CBD), after CHI, show a remarkable recovery profile in motor function compared to control untreated mice. Therefore, we propose to investigate the molecular basis of the neuroprotective role of CBD following CHI in mice. We will examine the optimal time course and dose of treatment with CBD after CHI. We will characterize changes in gene-expression in specific injured tissues in order to evaluate the molecular basis of CBD treatment. We will examine the possibility that part of the neuroprotection of CBD is due to epigenetic mechanism. To this end, we will analyze changes in gene methylations, histone modifications on chromatin structure and composition in mice treated with CBD in specific brain areas during the course of recovery. If successful, the results of the present study will provide the first evidence that the cannabis plant is useful for treating TBI and will set the basis for future deep examination of its neuroprotective potential. 

Hyperglycemia, inflammation and proximal tubular dysfunction contribute to the pathogenesis of diabetic nephropathy (DN), the leading cause of end-stage renal disease. Yet, the exact molecular signaling pathways underlying the development of DN are largely unknown. The endocannabinoid (eCB) system is critically involved in the onset of DN and globally-acting cannabinoid-1 (CB1) receptor antagonists were found effective in attenuating the development of DN and improving renal function. However, this treatment approach is no longer valid due to increasing centrally-mediated psychiatric side effects observed with rimonabant, the first-in-class commercially available CB1 inverse agonist.

My recent work has demonstrated that peripherally restricted CB1 antagonism may represent a novel approach to treat obesity and its related diabetic abnormalities without causing CNS-mediated side effects. Here, we propose to uncover the role of CB1 in the renal proximal tubular cells in the pathogenesis of DN, and the therapeutic potential of its blockade by peripherally restricted CB1 antagonist for the treatment of DN.

Our general hypothesis is that overactivation of the eCB system targets proximal tubular CB1 receptors to increase proteinuria, inflammation and nephropathy, leading to DN. The rationale for the proposed research is that understanding the contribution of the eCB system to the development of DN has the potential to translate into better understanding of the causes of the disease and identifying potential mechanisms that can be targeted for therapy.

 

Key words: Diabeteic nephropathy, CB1 receptor, Endocannabinoids, Renal dysfunction, Inflammation, Proteinuria

During the last few decades, there has been an epidemic increase worldwide in the prevalence of obesity and its metabolic and cardiovascular disorders. In addition, obesity has been increasingly recognized as an independent key factor in the development of renal complications, yet the underlying signaling pathways are not fully understood. Cannabinoid type-1 (CB1) receptor plays an important role in the onset of nephropathy associated with diabetes, and globally-acting CB1 receptor antagonists were found effective in improving renal function.

In this project, we propose to uncover the role of CB1, specifically in the renal proximal tubule, in obesity-induced renal dysfunction. Our general hypothesis is that increased activity of the renal endocannabinoid (eCB) system in obesity targets CB1 receptors in the renal proximal tubular cells (RPTCs) to promote RPTC dysfunction, and thus may contribute to the development of obesity-induced renal nephropathy.

To test this hypothesis, we will determine the metabolic and renal phenotypes associated with high-fat diet-induced obesity in a novel mouse strain that lacks CB1 receptors in the RPTCs. The rationale for the proposed research is that understanding the contribution of CB1/eCB system to the development of obesity-related kidney dysfunction has the potential to be translated into better understanding of the causes of the disease, and identifying potential mechanisms that can be targeted for therapy. This could support the development and clinical testing of pharmacological strategies, such as peripheral CB1 receptor antagonists for the treatment of not only obesity per-se but also to its related renal complications. 

 

Key words: Endocannabinoids, CB1 receptor, Obesity, Tubular inflammation & injury, Renal function, Renal fibrosis, Metabolism

The endocannabinoid (eCB) system is increasingly recognized as being of crucial importance in obesity-related metabolic abnormalities, one of which is liver injury and hepatic steatosis. eCBs, via activation of CB1 receptors in the liver, modulate hepatic lipogenesis and fatty acid oxidation. Yet, the underlying molecular mechanisms are largely unknown.

Our recent findings provide strong evidence for a functional role of the peroxisome proliferator-activated receptor-alpha (PPARα)/sirtuin 1(SIRT1) signaling pathway in potentially mediating the antisteatotic effect of peripherally restricted CB1 blockade. Therefore, the aim of this project is to delineate the key signaling factors involved in mediating the role of hepatic CB1 receptors in obesity-induced hepatic steatosis. Our general hypothesis is that activation of hepatic CB1 receptors by eCBs, decrease the expression and activity of SIRT1, which consequently impairs PPARα signaling, leading to the development of hepatic steatosis.

 

To achieve the objective of this project, we aim to: (i) verify the role of hepatic SIRT1 in the antisteatotic effect of peripheral CB1 blockade, (ii) examine the role of hepatic CB1 on SIRT1/PGC-1α/PPARα expression and activity, and (iii) determine the molecular signaling cascade by which CB1 modulates SIRT1 activity. As I have recently demonstrated the potential therapeutic value of selective blockade of peripheral CB1 receptors in animal models of obesity, further understanding of the regulation of hepatic steatosis by the CB1/eCB system has the potential to support the translation of our basic findings, and identify novel mechanisms that can be targeted for therapy, contributing directly to the fields of obesity and liver research.

Key words: Hepatic Steatosis, Liver injury, CB1 receptor blockade, PPAR alpha signaling, Sirt1, Obesity, Non-alcoholic fatty liver disease (NAFLD), Lipogenesis

Diabetes Research Unit, Institute for Drug Research, School of Pharmacy, Faculty of Medicine

Beta cells operate an hormetic mechanism to alleviate cytotoxic effects of free fatty acids. We have previously discovered that both high glucose and increasing palmitic acid levels induce substantial remodelling of membrane phospholipids and alter the relative abundance of saturated-, mono-unsaturated- and poly-unsaturated fatty acids. We aim at investigating the impact of the transformation of arachidonic acid into endocannabinoids under hyperglycemia and hyperlipidemic conditions. We have already reported that both conditions increase PLA2 activity and the hydrolysis of arachidonic acid from membrane phospholipids. Peroxidation of arachidonic acid leads to the generation of 4 hydroxynonenal that activates PPARδ and upregulates insulin secretion. Other studies assigned a regulatory role for endocannabinoids in beta cell function. We will investigate the mechanisms by which endocannabinoid biosynthesis is regulated in beta cells under nutrient overload stress.
 

Understanding the complex process of embryo implantation is a major goal of developmental biology. Such knowledge may also have important medical, social and economic implications, as 20-30% of all couples are temporarily (with 10% of them permanently) involuntarily childless. Of lost pregnancies, 75% are due to failure of implantation. Over the past several decades, it has become clear that shifts in different metabolic pathways determine the outcome of implantation, pointing to its regulation by a metabolic network.

 

Optimization of the metabolic environment to be favorable for embryo implantation is currently at the core of assisted conception programmes. However, despite intensive research, the pathways that are involved in implantation, and how they interact, are still poorly understood. Although no systemic overview for metabolic processes during implantation has been offered thus far, specific metabolic pathways have been suggested to be involved in embryo implantation including ovarian hormones signaling pathways, central carbon metabolism pathways, inflammatory signaling as well as lysophosphatidic acid signaling.

Importantly, a large volume of literature established an important role for the endocannabinoid system in the implantation of the mammalian embryo.

Advances in analytical chemistry and in bio-informatic software provide a means for the unbiased analysis of the metabolic processes by metabolomics: the profiling of the metabolome. The metabolome represents the collection of all metabolites in a biological sample. A systematic study of the unique chemical fingerprints that are the result of specific cellular processes will shed important new light on the process of embryo implantation.

­­­We are looking into the metabolism and specifically into endocannabinoid metabolism of the pre-implanted embryo using medium from In Vitro Fertilization of human embryos. We are also utilizing stem cell technologies to study the metabolic maturation of the implanted embryo, and developed a multi-cellular model for the examination of cellular metabolic interactions during implantation.  

Key words: Metabolomics; embryo implantation; embryonic stem cells; trophectoderm; endometrium

The effect of endocannabinoid receptors on receptiveness of the endometrium

Plasma AEA levels fluctuate with the natural menstrual cycle, with the highest levels during the follicular phase. Likewise, the expression and distribution of FAAH, NAPE-PLD, CB1 and CB2 in female reproductive tissues also vary with the menstrual cycle suggesting that the eCS expression is under hormonal control.

Implantation is the very early stage of pregnancy at which the blastocyst adheres and invades the wall of the uterus. The process of implantation implies a highly synchronized and complex bidirectional communication between the conceptus and the luminal endometrial cells, and it could be divided in three stages: apposition, adhesion and penetration.

The endocannabinoid system plays an important role during the process of implantation. Studies in mice have shown that a delicate balance between anandamide (AEA) synthesis (driven mainly by NAPE-PLD) and degradation (mainly by FAAH) is necessary to ensure an appropriate ‘AEA tone’ during implantation.

In our studies, we examine the role of the endocannabinoid receptors in patients with repeated implantation failure (RIF).

 

Are endocannabinoids involved in the creation of metastatic niche?

 Endocannabinoids are produced in several peripheral tissues resulting in cell-type and location-specific profiles so that the eCB pattern in the tumor microenvironment depends on the tumor’s origin and site of primary growth and metastasis. Tumor growth is associated with an increase of 2-arachidonoylglycerol (2-AG) both at the site of the primary tumor and in plasma of the patients. It steadily increased over the course of cancer development and metastasis, suggesting that the growing tumor and circulating metastatic tumor cells secrete large amounts of 2-AG, sufficient enough to manifest in high plasma concentrations.

It is likely that the tumor itself mainly secretes 2-AG. The endocannabinoids have diverse effects on cannabinoid receptors including the typical cannabinoid receptor 1 and 2 (CB1, CB2) and other non typical ones.

Treatment with exo- and endogenous cannabinoids reduced cancer growth in several rodent models and cell culture experiments, but CB2 expression of the tumor itself has been recently associated with poor prognosis in breast cancer.

In our studies we examine the effect of the endocannabinoid system on the early stages of metastatic spread, the formation of the metastatic niche.

 

Key words: ovarian cancer , exosomes, ncRNAs, tumor progression

 

Introduction: The activity of the cannabis plant extracts, synthetic and endo-cannabinoids derivatives is well documented in eukaryotic systems. However, very few studies have been published on the effect of cannabinoids on microorganism (e.g. bacteria or fungi) and none on their effect on biofilm formation or on microbe's cell-cell communication (quorum sensing). 

Biofilm is the most abundant state in which microbes are found in nature. Most infectious diseases are associated with biofilms. Quorum sensing has been shown to regulate virulence factors pathogenesis, motility and biofilm formation.

 

Innovative of the research: The aim of our research group is to investigate the effect of plant, synthetic and endo-cannabinoids on microorganisms.

Significance of the research: An insight into the mechanism of action of cannabis cannabinoids and endo-cannabinoids on microbes and open new understanding of potential cannabinoid-microbes interaction, which was hardly investigated.

More so, the exciting possibility that endo cannabinoids may have also present endogenous anti biofilm/anti quorum sensing activity is being examined. This will indicate a new line of defense of the human against bacterial invasion.

Administration of cannabinoids (that affect biofilm) together with antimicrobial agents may in the future open the door to \a possible tool to overcome bacterial resistance to antibiotics.

Methods used in the laboratory: The effect of cannabinoids and endo-cannabinoids are examined in batch and flows biofilm models. Biofilm structure, metabolic activity, in silico analysis, motility, gene expression are among the techniques applied in the laboratory.

 

Results: Analysis of quorum sensing in wild-type and mutant strains of V. harveyi revealed that a cannabinoid derivative (HU-210) affects the quorum sensing cascade via auto inducer-2 (AI-2) pathway. Furthermore, this effect was correlated to biofilm formation and swimming motility of the bacteria that were significantly reduced in the presence of HU-210. HU-210 also altered the expression of genes which are associated with quorum sensing, specifically downregulating the genes of the AI-2 quorum sensing cascade. Preliminary results have also indicated that endo-cannabinoid (anandamide-ANA) has also an effect of bacteria at concentration less than MIC.

Conclusions: Are the cannabinoids the new era in repacking antibiotics and anti-fungal agents? According to our results, some cannabinoids derivatives do have an antibiofilm effect.

First evidence is provided in our study for interference of bacterial signal-transduction systems by a synthetic cannabinoid. Its effect via the AI-2 cascade (universal AI) opens a vast possibility of new mechanisms of action that have not been explored before. More so, our results opens new pharmaceutical avenues for the commercial use of derivative of cannabinoids for various, human related and non-human biofilms,

Key words: Cannabis, cannabinoids, biofilm, quorum sensing, microorganisms