Effects of CB2 and TRPV1 receptors’ stimulation in pediatric acute T-lymphoblastic leukemia

Abstract

T-Acute Lymphoblastic Leukemia (T-ALL) is less frequent than B-ALL, but it has poorer outcome. For this reason new therapeutic approaches are needed to treat this malignancy.The Endocannabinoid/Endovanilloid (EC/EV) system has been proposed as possible target to treat several malignancies, including lymphoblastic diseases. The EC/EV system is composed of two G-Protein Coupled Receptors (CB1 and CB2), the Transient Potential Vanilloid 1 (TRPV1) channel, their endogenous and exogenous ligands and enzymes. CB1 is expressed mainly in central nervous system while CB2 predominantly on immune and peripheral cells, therefore we chose to selectively stimulate CB2 and TRPV1.We treated T-ALL lymphoblasts derived from 4 patients and Jurkat cells with a selective agonist at CB2 receptor: JWH-133 [100 nM] and an agonist at TRPV1 calcium channel: RTX [5 uM] at 6, 12 and 24 hours. We analyzed the effect on apoptosis and Cell Cycle Progression by a cytofluorimetric assays and evaluated the expression level of several target genes (Caspase 3, Bax, Bcl-2, AKT, ERK, PTEN, Notch-1, CDK2, p53) involved in cell survival and apoptosis, by Real-Time PCR and Western Blotting.We observed a pro-apoptotic, anti-proliferative effect of these compounds in both primary lymphoblasts obtained from patients with T-ALL and in Jurkat cell line. Our results show that both CB2 stimulation and TRPV1 activation, can increase the apoptosis in vitro, interfere with cell cycle progression and reduce cell proliferation, indicating that a new therapeutic approach to T-cell ALL might be possible by modulating CB2 and TRPV1 receptors.

Pharmacological Properties, Therapeutic Potential and Molecular Mechanisms of JWH133, a CB2 Receptor-Selective Agonist

“…Leukemias account for 30% of all pediatric cancers, and acute lymphoblastic leukemia (ALL) is the most prevalent pediatric leukemia, representing 75% of all pediatric leukemia cases (Terwilliger and Abdul-Hay, 2017). Punzo et al (2018a) showed that JWH133 promoted apoptosis in patients with T-ALL and a Jurkat cell line via enhanced caspase-3 expression and Bax/Bcl-2 ratio. Moreover, JWH133 prevented tumor cell growth and survival via reduced expression of AKT, ERK, and Notch-1, while increasing the expression of PTEN and p53.…”

Transient Receptor Potential (TRP) Channels in Haematological Malignancies: An Update

“…The activation of TRPV1 channels by resiniferatoxin (RTX), a potent analogue of CPS and a TRPV agonist, at 5 µM, arrests the cell cycle at G0/G1 phase, reduces cell proliferation, and induces apoptosis in both Jurkat cells and cellderived T-ALL patients. In particular, after 6-12 h of treatment, a reduction of AKT, ERK, Notch-1, CDK2, and phospho-CDK2 protein levels, together with PTEN and p53 proteins upregulation in RTX-treated cells, has been reported [37]. Supporting data about the antioncogenic activity of TRPV1 were disclosed by Cetintas et al In this study, CPS inhibits the proliferation of CCRF-CEM cells in a dose-dependent manner, inducing apoptosis via caspase-3 activation and reduction of Bcl-2 expression.…”

“…For instance, the TRPC3 channel blocker, Pyr3, enhances apoptosis induced by dexamethasone in ALL cells isolated from patients by altering calcium signalling, mitochondrial membrane potential, and ROS production [44]. In addition, the activation of TRPV1, by using the specific agonist RTX, reduced cell proliferation, blocked cell cycle, and increased apoptosis in T cells from ALL patients [37].…”

Hirsutanol A inhibits T‑acute lymphocytic leukemia Jurkat cell viability through cell cycle arrest and p53‑dependent induction of apoptosis

“…T-ALL is caused by the diffuse bone marrow infiltration of immature T lymphoblasts, which clinically manifests as central nervous system infiltration and mediastinal mass with pleural effusion in the early stage (6). T-ALL is far more difficult to treat than B-ALL and has a poor long-term prognosis (7,8) resulting from poor sensitivity and resistance to chemotherapeutic drugs, issues with remission and clearance of bone marrow microresidual lesions, and a high recurrence rate in the central nervous system (9,10). With the advancement of chemotherapy regimens, the 5-year disease-free survival rate of pediatric T-ALL has substantially improved (11)(12)(13).…”