Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission
Transcription factor programming of pluripotent stem cells (PSCs) has emerged as an approach to generate human neurons for disease modeling. However, programming schemes produce a variety of cell types, and those neurons that are made often retain an immature phenotype, which limits their utility in...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Published: |
Elsevier
2018
|
| Online Access: | http://hdl.handle.net/1721.1/117674 https://orcid.org/0000-0002-8021-277X |
| _version_ | 1811070199755964416 |
|---|---|
| author | Nehme, Ralda Zuccaro, Emanuela Ghosh, Sulagna Dia Li, Chenchen Sherwood, John L. Pietilainen, Olli Barrett, Lindy E. Limone, Francesco Worringer, Kathleen A. Kommineni, Sravya Zang, Ying Cacchiarelli, Davide Meissner, Alex Adolfsson, Rolf Haggarty, Stephen Madison, Jon Muller, Matthias Arlotta, Paola Fu, Zhanyan Feng, Guoping Eggan, Kevin |
| author2 | Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences |
| author_facet | Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Nehme, Ralda Zuccaro, Emanuela Ghosh, Sulagna Dia Li, Chenchen Sherwood, John L. Pietilainen, Olli Barrett, Lindy E. Limone, Francesco Worringer, Kathleen A. Kommineni, Sravya Zang, Ying Cacchiarelli, Davide Meissner, Alex Adolfsson, Rolf Haggarty, Stephen Madison, Jon Muller, Matthias Arlotta, Paola Fu, Zhanyan Feng, Guoping Eggan, Kevin |
| author_sort | Nehme, Ralda |
| collection | MIT |
| description | Transcription factor programming of pluripotent stem cells (PSCs) has emerged as an approach to generate human neurons for disease modeling. However, programming schemes produce a variety of cell types, and those neurons that are made often retain an immature phenotype, which limits their utility in modeling neuronal processes, including synaptic transmission. We report that combining NGN2 programming with SMAD and WNT inhibition generates human patterned induced neurons (hpiNs). Single-cell analyses showed that hpiN cultures contained cells along a developmental continuum, ranging from poorly differentiated neuronal progenitors to well-differentiated, excitatory glutamatergic neurons. The most differentiated neurons could be identified using a CAMK2A::GFP reporter gene and exhibited greater functionality, including NMDAR-mediated synaptic transmission. We conclude that utilizing single-cell and reporter gene approaches for selecting successfully programmed cells for study will greatly enhance the utility of hpiNs and other programmed neuronal populations in the modeling of nervous system disorders. Nehme et al. combine two strong neuralizing factors (transcription factor programming and small molecule patterning) to generate human excitatory neurons from stem cells. They further undertake single-cell and reporter gene approaches to select highly differentiated neurons with increased functionality, augmenting their utility in the modeling of nervous system disorders. |
| first_indexed | 2024-09-23T08:31:44Z |
| format | Article |
| id | mit-1721.1/117674 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T08:31:44Z |
| publishDate | 2018 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1176742022-09-30T09:28:03Z Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission Nehme, Ralda Zuccaro, Emanuela Ghosh, Sulagna Dia Li, Chenchen Sherwood, John L. Pietilainen, Olli Barrett, Lindy E. Limone, Francesco Worringer, Kathleen A. Kommineni, Sravya Zang, Ying Cacchiarelli, Davide Meissner, Alex Adolfsson, Rolf Haggarty, Stephen Madison, Jon Muller, Matthias Arlotta, Paola Fu, Zhanyan Feng, Guoping Eggan, Kevin Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Feng, Guoping Transcription factor programming of pluripotent stem cells (PSCs) has emerged as an approach to generate human neurons for disease modeling. However, programming schemes produce a variety of cell types, and those neurons that are made often retain an immature phenotype, which limits their utility in modeling neuronal processes, including synaptic transmission. We report that combining NGN2 programming with SMAD and WNT inhibition generates human patterned induced neurons (hpiNs). Single-cell analyses showed that hpiN cultures contained cells along a developmental continuum, ranging from poorly differentiated neuronal progenitors to well-differentiated, excitatory glutamatergic neurons. The most differentiated neurons could be identified using a CAMK2A::GFP reporter gene and exhibited greater functionality, including NMDAR-mediated synaptic transmission. We conclude that utilizing single-cell and reporter gene approaches for selecting successfully programmed cells for study will greatly enhance the utility of hpiNs and other programmed neuronal populations in the modeling of nervous system disorders. Nehme et al. combine two strong neuralizing factors (transcription factor programming and small molecule patterning) to generate human excitatory neurons from stem cells. They further undertake single-cell and reporter gene approaches to select highly differentiated neurons with increased functionality, augmenting their utility in the modeling of nervous system disorders. 2018-09-07T17:23:53Z 2018-09-07T17:23:53Z 2018-05 2018-03 2018-08-30T12:48:34Z Article http://purl.org/eprint/type/JournalArticle 2211-1247 http://hdl.handle.net/1721.1/117674 Nehme, Ralda, et al. “Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission.” Cell Reports, vol. 23, no. 8, May 2018, pp. 2509–23. © 2018 The Authors https://orcid.org/0000-0002-8021-277X http://dx.doi.org/10.1016/j.celrep.2018.04.066 Cell Reports Creative Commons Attribution 4.0 International License http://creativecommons.org/licenses/by/4.0/ application/pdf Elsevier Elsevier |
| spellingShingle | Nehme, Ralda Zuccaro, Emanuela Ghosh, Sulagna Dia Li, Chenchen Sherwood, John L. Pietilainen, Olli Barrett, Lindy E. Limone, Francesco Worringer, Kathleen A. Kommineni, Sravya Zang, Ying Cacchiarelli, Davide Meissner, Alex Adolfsson, Rolf Haggarty, Stephen Madison, Jon Muller, Matthias Arlotta, Paola Fu, Zhanyan Feng, Guoping Eggan, Kevin Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission |
| title | Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission |
| title_full | Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission |
| title_fullStr | Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission |
| title_full_unstemmed | Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission |
| title_short | Combining NGN2 Programming with Developmental Patterning Generates Human Excitatory Neurons with NMDAR-Mediated Synaptic Transmission |
| title_sort | combining ngn2 programming with developmental patterning generates human excitatory neurons with nmdar mediated synaptic transmission |
| url | http://hdl.handle.net/1721.1/117674 https://orcid.org/0000-0002-8021-277X |
| work_keys_str_mv | AT nehmeralda combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT zuccaroemanuela combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT ghoshsulagnadia combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT lichenchen combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT sherwoodjohnl combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT pietilainenolli combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT barrettlindye combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT limonefrancesco combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT worringerkathleena combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT komminenisravya combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT zangying combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT cacchiarellidavide combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT meissneralex combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT adolfssonrolf combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT haggartystephen combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT madisonjon combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT mullermatthias combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT arlottapaola combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT fuzhanyan combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT fengguoping combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission AT eggankevin combiningngn2programmingwithdevelopmentalpatterninggenerateshumanexcitatoryneuronswithnmdarmediatedsynaptictransmission |