Background & Aims
Dorsal root ganglia (DRG) contain neuronal soma that receive sensory input from peripheral organs and transmit it to the central nervous system. These neurons are thus the first point of integration of sensory signals from the body’s internal and external environment. Sensory neurons can be affected by dysimmune, toxic, metabolic, infectious, hereditary, and idiopathic ganglionopathies, resulting in injury and chronic pain. Recurrent pain is the most prevalent form of neuropathy, affecting nearly 20% of people worldwide, with only very few new treatments in the last decade. Sensory neuron activity is modulated by the glial and immune microenvironment in the DRG. Satellite glial cells (SGCs) surround neuronal soma and represent an essential component of peripheral pain transmission. Although rodent model systems have been very useful for understanding the basic mechanisms of pain perception, very little cellular and molecular information is available on human DRGs (hDRGs) (1,2).
Methods
A better understanding of the role of the DRG microenvironment is necessary to understand how peripheral neurons process pain in human. To determine the molecular and spatial properties of individual cells within hDRG, we combined ATAC-seq and RNAseq (snMultiome) (3) with imaging mass cytometry (IMC) (4) as the two main profiling methods for unbiased gene and protein assessment.
Results
We developed a nuclear purification protocol for snMultiome to enrich for neurons, since neurons account for only 1.3% of all cells in hDRG. We also customized an antibody panel to profile neurons, glia and immune cells in hDRG by IMC.
Conclusions
Applying this unique spatially resolved, highly-multiplexed nuclear transcriptomic and proteomic approach to pain research is transforming our ability to interrogate changes in cell populations and gene expression in human DRG.
References
1. Mogil, J.S. Animal models of pain: progress and challenges. Nat Rev Neurosci 10, 283-294 (2009).
2. Vierck, C.J., Hansson, P.T. & Yezierski, R.P. Clinical and pre-clinical pain assessment: are wemeasuring the same thing? Pain 135, 7-10 (2008).
3. Butler, A., Hoffman, P., Smibert, P., Papalexi, E. & Satija, R. Integrating single-cell transcriptomic data
across different conditions, technologies, and species. Nature biotechnology 36, 411-420 (2018).
4. Baharlou, H., Canete, N.P., Cunningham, A.L., Harman, A.N. & Patrick, E. Mass Cytometry Imaging for the Study of Human Diseases-Applications and Data Analysis Strategies. Front Immunol 10, 2657 (2019).
Presenting Author
Pauline Meriau
Poster Authors
Pauline Meriau
PhD
Washington University in St.Louis
Lead Author
Kevin Boyer
B.S.
Washington University School of Medicine, St. Louis, MO
Lead Author
Huma Naz
PhD
Washington University School of Medicine, St. Louis, MO
Lead Author
Ibrahim Ibrahim Olabayode Saliu
PhD
Washington University School of Medicine, St. Louis, MO
Lead Author
Maria Payne
Washington University School of Medicine, St. Louis, MO
Lead Author
Lite Yang
Washington University School of Medicine, St. Louis, MO
Lead Author
John Del Rosario
PhD
Washington University School of Medicine, St. Louis, MO
Lead Author
Adam Dourson
Washington University
Lead Author
Alexander Chamessian
MD,PhD
Washington University School of Medicine, St. Louis, MO
Lead Author
Jun-Nan Li
Washington University School of Medicine, St. Louis, MO
Lead Author
Jiwon Yi
Washington University School of Medicine, St. Louis, MO
Lead Author
Bryan A Copits
PhD
Washington University School of Medicine, St. Louis, MO
Lead Author
Robert W Gereau
Pr
Washington University School of Medicine, St. Louis, MO
Lead Author
Guoyan Zhao
Washington University School of Medicine, St. Louis, MO
Lead Author
Topics
- Mechanisms: Biological-Systems (Physiology/Anatomy)