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)