Design and biological assessment of membrane-tethering neuroprotective peptides derived from the pituitary adenylate cyclase-activating polypeptide type 1 receptor

• Published in: Biochimica et biophysica acta. General subjects Vol. 1863; no. 11; p. 129398
• Main Authors: Poujol de Molliens, Mathilde, Jamadagni, Priyanka, Létourneau, Myriam, Devost, Dominic, Hébert, Terence E., Patten, Shunmoogum A., Fournier, Alain, Chatenet, David
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.11.2019
• Subjects: Animals; BRET-based biosensor; Cell Line, Tumor; Cell Survival - drug effects; HEK293 Cells; Humans; Neurogenesis - drug effects; Neuroprotection; Neuroprotective Agents - chemistry; Neuroprotective Agents - pharmacology; PAC1 receptor; PACAP; Parkinson's disease; Pepducin; Peptides - chemistry; Peptides - pharmacology; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I - chemistry; Zebrafish - embryology; Neuroprotection; DIEA; Parkinson's disease; BBB; DMEM; P/S; FBS; Boc; BRET-based biosensor; Fmoc; PAC1; dpf; DMF; RlucII; hpf; BRET; BOP; MO; PACAP; MALDI-TOF; GFP 10; ACN; PAC1 receptor; DCM; RP-HPLC; vDC; VPAC; Pepducin; GPCR; Tel; VIP
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/j.bbagen.2019.07.007

The pituitary adenylate cyclase-activating polypeptide (PACAP) type 1 receptor (PAC1), a class B G protein-coupled receptor (GPCR), has emerged as a promising target for treating neurodegenerative conditions. Unfortunately, despite years of research, no PAC1-specific agonist has been discovered, as activity on two other GPCRs, VPAC1 and VPAC2, is retained with current analogs. Cell signaling is related to structural modifications in the intracellular loops (ICLs) of GPCRs. Thus, we hypothesized that peptides derived from the ICLs (called pepducins) of PAC1 might initiate, as allosteric ligands, signaling cascades after recognition of the parent receptor and modulation of its conformational landscape. Three pepducins were synthesized and evaluated for their ability to 1) promote cell survival; 2) stimulate various signaling pathways associated with PAC1 activation; 3) modulate selectively PAC1, VPAC1 or VPAC2 activation; and 4) sustain mobility and prevent death of dopaminergic neurons in a zebrafish model of neurodegeneration. Assays demonstrated that these molecules promote SH-SY5Y cell survival, a human neuroblastoma cell line expressing PAC1, and activate signaling via Gαs and Gαq, with distinct potencies and efficacies. Also, PAC1-Pep1 and PAC1-Pep2 activated selectively PAC1-mediated Gαs stimulation. Finally, experiments, using a zebrafish neurodegeneration model, showed a neuroprotective action with all three pepducins and in particular, revealed the ability of PAC1-Pep1 and PAC1-Pep3 to preserve fish mobility and tyrosine hydroxylase expression in the brain. We have developed the first neuroprotective pepducins derived from PAC1, a class B GPCR. PAC1-derived pepducins represent attractive templates for the development of innovative neuroprotecting molecules. •PAC1-derived pepducins can promote SH-SY5Y cell survival.•PAC1-Pep1 and PAC1-Pep2 can selectively trigger PAC1-mediated Gas stimulation.•PAC1-derived pepducins preserve fish mobility and tyrosine hydroxylase expression.