Content area
Full Text
A R T I C L E S
Modeling pain in vitro using nociceptor neurons reprogrammed from fibroblasts
Brian J Wainger13,8, Elizabeth D Buttermore1,3,8, Julia T Oliveira1,4, Cassidy Mellin1, Seungkyu Lee1,3, Wardiya Afshar Saber1, Amy J Wang1, Justin K Ichida5,6, Isaac M Chiu1,3, Lee Barrett1, Eric A Huebner1,3, Canan Bilgin1, Naomi Tsujimoto5, Christian Brenneis1, Kush Kapur1, Lee L Rubin5, Kevin Eggan5,7 & Clifford J Woolf1,3
Reprogramming somatic cells from one cell fate to another can generate specific neurons suitable for disease modeling.
To maximize the utility of patient-derived neurons, they must model not only disease-relevant cell classes, but also the diversity of neuronal subtypes found in vivo and the pathophysiological changes that underlie specific clinical diseases. We identified five transcription factors that reprogram mouse and human fibroblasts into noxious stimulusdetecting (nociceptor) neurons. These recapitulated the expression of quintessential nociceptor-specific functional receptors and channels found in adult mouse nociceptor neurons, as well as native subtype diversity. Moreover, the derived nociceptor neurons exhibited TrpV1 sensitization to the inflammatory mediator prostaglandin E2 and the chemotherapeutic drug oxaliplatin, modeling the inherent mechanisms underlying inflammatory pain hypersensitivity and painful chemotherapy-induced neuropathy. Using fibroblasts from patients with familial dysautonomia (hereditary sensory and autonomic neuropathy type III), we found that the technique was able to reveal previously unknown aspects of human disease phenotypes in vitro.
npg 201 5 Nature America, Inc. All rights reserved.
Directed differentiation from pluripotent stem cells and lineage reprogramming of fibroblasts can both be used to derive a wide range of different neuronal subtypes1,2. Although the known sequence of morphogen exposure and consequent molecular changes in the development of specific neurons can guide directed differentiation strategies, the selection of transcription factors for lineage reprogramming from fibroblasts remains essentially empirical. No single transcription factor has proven to be essential for driving cell fates in all of the neuronal reprogramming studies to date, despite the fact that specific factors such as Ascl1 or Ngn2 seem particularly potent in deriving a range of different neuronal subtypes3. Brn2, Ascl1 and Myt1l (or BAM) generate generic neurons on their own4 and specific neuronal subtypes when combined with additional factors5. Moreover, the developmental stage at which a particular transcription factor acts in vivo may determine whether that factor facilitates or inhibits the patterning of...