ABSTRACT
ABSTRACT: Individuals living with sickle cell disease (SCD) experience severe recurrent acute and chronic pain. Challenges to gaining mechanistic insight into pathogenic SCD pain processes include differential gene expression and function of sensory neurons between humans and mice with SCD, and extremely limited availability of neuronal tissues from patients with SCD. Here, we used induced pluripotent stem cells (iPSCs), derived from patients with SCD, differentiated into sensory neurons (SCD iSNs) to begin to overcome these challenges. We characterize key gene expression and function of SCD iSNs to establish a model to investigate intrinsic and extrinsic factors that may contribute to SCD pain. Despite similarities in receptor gene expression, SCD iSNs show pronounced excitability using patch clamp electrophysiology. Furthermore, we find that plasma taken from patients with SCD during acute pain associated with a vaso-occlusive event increases the calcium responses to the nociceptive stimulus capsaicin in SCD iSNs compared with those treated with paired plasma from patients with SCD at steady state baseline or healthy control plasma samples. We identified high levels of the polyamine spermine in baseline and acute pain states of plasma from patients with SCD, which sensitizes SCD iSNs to subthreshold concentrations of capsaicin. Together, these data identify potential intrinsic mechanisms within SCD iSNs that may extend beyond a blood-based pathology.
Subject(s)
Anemia, Sickle Cell , Induced Pluripotent Stem Cells , Sensory Receptor Cells , Humans , Anemia, Sickle Cell/blood , Anemia, Sickle Cell/pathology , Induced Pluripotent Stem Cells/metabolism , Induced Pluripotent Stem Cells/cytology , Sensory Receptor Cells/metabolism , Sensory Receptor Cells/physiology , Sensory Receptor Cells/pathology , Cell Differentiation , Capsaicin/pharmacology , Male , Female , Plasma/metabolismABSTRACT
Mutations in ASAH1 have been linked to two allegedly distinct disorders: Farber disease (FD) and spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME). We have previously reported FD-like phenotypes in mice harboring a single amino acid substitution in acid ceramidase (ACDase), P361R, known to be pathogenic in humans (P361R-Farber). Here we describe a mouse model with an SMA-PME-like phenotype (P361R-SMA). P361R-SMA mice live 2-3-times longer than P361R-Farber mice and have different phenotypes including progressive ataxia and bladder dysfunction, which suggests neurological dysfunction. We found profound demyelination, loss of axons, and altered sphingolipid levels in P361R-SMA spinal cords; severe pathology was restricted to the white matter. Our model can serve as a tool to study the pathological effects of ACDase deficiency on the central nervous system and to evaluate potential therapies for SMA-PME.
Subject(s)
Farber Lipogranulomatosis , Muscular Atrophy, Spinal , Myoclonic Epilepsies, Progressive , Humans , Mice , Animals , Farber Lipogranulomatosis/genetics , Farber Lipogranulomatosis/metabolism , Farber Lipogranulomatosis/pathology , Muscular Atrophy, Spinal/genetics , Muscular Atrophy, Spinal/pathology , Sphingolipids/metabolism , Myoclonic Epilepsies, Progressive/genetics , Myoclonic Epilepsies, Progressive/pathology , PhenotypeABSTRACT
Epidermal keratinocytes mediate touch sensation by detecting and encoding tactile information to sensory neurons. However, the specific mechanotransducers that enable keratinocytes to respond to mechanical stimulation are unknown. Here, we found that the mechanically-gated ion channel PIEZO1 is a key keratinocyte mechanotransducer. Keratinocyte expression of PIEZO1 is critical for normal sensory afferent firing and behavioral responses to mechanical stimuli in mice.
