Delta-9-tetrahydrocannabinol inhibits invasion of HTR8/SVneo human extravillous trophoblast cells and negatively impacts mitochondrial function

Abstract

Prenatal cannabis use is a significant problem and poses important health risks for the developing fetus. The molecular mechanisms underlying these changes are not fully elucidated but are thought to be attributed to delta-9-tetrahydrocannabinol (THC), the main bioactive constituent of cannabis. It has been reported that THC may target the mitochondria in several tissue types, including placental tissue and trophoblast cell lines, and alter their function. In the present study, in response to 48-h THC treatment of the human extravillous trophoblast cell line HTR8/SVneo, we demonstrate that cell proliferation and invasion are significantly reduced. We further demonstrate THC-treatment elevated levels of cellular reactive oxygen species and markers of lipid damage. This was accompanied by evidence of increased mitochondrial fission. We also observed increased expression of cellular stress markers, HSP70 and HSP60, following exposure to THC. These effects were coincident with reduced mitochondrial respiratory function and a decrease in mitochondrial membrane potential. Taken together, our results suggest that THC can induce mitochondrial dysfunction and reduce trophoblast invasion; outcomes that have been previously linked to poor placentation. We also demonstrate that these changes in HTR8/SVneo biology may be variably mediated by cannabinoid receptors CB1 and CB2.

Glucocorticoids, Stress and Delta-9 Tetrahydrocannabinol (THC) during Early Embryonic Development

“…These observations provide evidence to suggest that genes encoding for HSP70 may be a common downstream target for both glucocorticoid-and THC-mediated signaling. In contrast to speculations derived from the studies of Natale et al (2020), Walker et al (2021) suggested that THC does not buffer subcellular stress signaling through interactions with CB1 and CB2 within the human trophectoderm [24]. They demonstrated that exposure to 20 µM THC significantly increased expression of HSPs 60 and 70, but not antioxidant enzyme mRNA within HTR8/SVneo cells.…”

“…The successful production and subsequent development of an embryo from competent oocytes is dependent on many factors that are influenced by the ECS. Namely, CB1 receptors have been localized on the outer mitochondrial membrane, suggesting that the ECS regulates the utilization of cellular energy and apoptosis [24,[33][34][35]. The direct roles of the ECS during embryonic development is yet to be fully elucidated; however, multiple studies have suggested that the ECS influences embryo development directly and indirectly by modulating gene expression and mitochondrial processes related to apoptosis and cellular energy balance [23,24,[33][34][35][36].…”

“…The hormonal microenvironment is tightly regulated throughout oocyte maturation, given that oocyte competence, quality, and capacity to produce an embryo can be altered by disruption to such processes [20][21][22]. Given the capacity of both the ECS and glucocorticoids to alter reproductive processes [12,15,23,24,31,34,49,50,54,56,57], it is important to investigate potential interactions between these molecules and their signaling pathways within reproductive tissues. Both glucocorticoids and THC have been shown to influence multiple signaling pathways involved in oocyte and embryo development including extracellular signal-regulated kinase (ERK1/2), cyclic AMP/protein kinase A (cAMP/PKA), Protein kinase B (PKB)/AKT, and mitogen-activated protein kinases (MAPK) p38 [12][13][14]30,58] (Figure 1).…”

“…Additionally, HSP70 is the most conserved chaperone across all mammalian species, thus, results obtained from animal models is largely applicable to humans [105]. Both THC (20 µM) and Cortisol (0.1 µg/mL) exposure have been shown to increase the expression of HSP70 in human trophoblasts and bovine embryos, respectively [24,54]. These observations provide evidence to suggest that genes encoding for HSP70 may be a common downstream target for both glucocorticoid-and THC-mediated signaling.…”

“…However, there is a lack of research that explores the potential interactions between signaling pathways activated by THC and stress hormones at the level of the oocyte and embryo during early development. In addition, there have been no reports of THC-induced teratogenic effects, irrespective of its ability to cross the placental barrier [24]. Strong correlations between rising cannabis use and stress-related symptoms among women of reproductive age suggests that there is a lack of accessible research available for individuals to make more informed decisions regarding their reproductive health and for physicians to properly guide their patients.…”