ABSTRACT
Pulmonary arterial hypertension (PAH) has demonstrated multi-serotonin receptor dependent pathologies, characterized by increased tone (5-HT1B receptor) and complex lesions (SERT, 5-HT1B, 5-HT2B receptors) of the pulmonary vasculature together with right ventricular hypertrophy, ischemia and fibrosis (5-HT2B receptor). Selective inhibitors of individual signaling elements - SERT, 5-HT2A, 5HT2B, and combined 5-HT2A/B receptors, have all been tested clinically and failed. Thus, inhibition of tryptophan hydroxylase 1 (TPH1), the rate limiting step in 5-HT synthesis, has been suggested as a more broad, and thereby more effective, mode of 5-HT inhibition. However, selectivity over non-pathogenic enzyme family members, TPH2, phenylalanine hydroxylase, and tyrosine hydroxylase has hampered therapeutic development. Here we describe the site/sequence, biochemical, and biophysical characterization of a novel allosteric site on TPH1 through which selectivity over TPH2 and related aromatic amino acid hydroxylases is achieved. We demonstrate the mechanism of action by which novel compounds selectively inhibit TPH1 using surface plasma resonance and enzyme competition assays with both tryptophan ligand and BH4 co-factor. We demonstrate 15-fold greater potency within a human carcinoid cell line versus the most potent known TPH1/2 non-specific inhibitor. Lastly, we detail a novel canine in vivo system utilized to determine effective biologic inhibition of newly synthesized 5-HT. These findings are the first to demonstrate TPH1-selective inhibition and may pave the way to a truly effective means to reduce pathologic 5-HT and thereby treat complex remodeling diseases such as PAH.
ABSTRACT
The accessory nerve is traditionally described as having both spinal and cranial roots, with the spinal root originating from the upper cervical segments of the spinal cord and the cranial root originating from the dorsolateral surface of the medulla oblongata. The spinal rootlets and cranial rootlets converge either before entering the jugular foramen or within it. In a recent report, this conventional view has been challenged by finding no cranial contribution to the accessory nerve. The present study was undertaken to re-examine the accessory and vagus nerves within the cranium and jugular foramen, with particular emphasis on the components of the accessory nerve. These nerves were traced from their rootlets attaching to the spinal cord and the medulla and then through the jugular foramen. The jugular foramen was exposed by removing the dural covering and surrounding bone. A surgical dissecting microscope was used to trace the roots of the glossopharyngeal nerve (CN IX), vagus nerve (CN X) and accessory nerve (CN XI) before they entered the jugular foramen and during their travel through it. The present study demonstrates that the accessory nerve exists in two forms within the cranial cavity. In the majority of cases (11 of 12), CN XI originated from the spinal cord with no distinct contribution from the medulla. However, in one of 12 cases, a small but distinct connection was seen between the vagus and the spinal accessory nerves within the jugular foramen.