Inter- and intracellular resistance and vulnerability in motor neuron diseases
Author: Nijssen, Jik
Date: 2020-05-27
Location: Eva and Georg Klein, Solnavägen 9, Biomedicum, Karolinska Institutet, Solna
Time: 09.30
Department: Inst för neurovetenskap / Dept of Neuroscience
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Thesis (13.67Mb)
Abstract
Amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA) are fatal diseases
presenting with degeneration and loss of lower motor neurons in the brainstem and spinal cord.
However, oculomotor and trochlear motor neurons that control eye movement, are resistant
until late stages of disease. Moreover, individual motor neurons degenerate in a distinct pattern.
Firstly, the distal synapse between motor neuron and muscle, the neuromuscular junction
(NMJ), becomes denervated. Subsequently, the neuron degenerates in a retrograde manner,
until the cell body in the spinal cord is lost later in disease. This highlights the distal axon and
NMJ as the most vulnerable subcellular compartment of motor neurons.
We set out to investigate the mechanisms that underlie the resistance of oculomotor neurons, as well as the differential vulnerability within motor neurons themselves. We first validated the resistance of oculomotor neurons in two mouse models of ALS and SMA in Paper I. Next, we performed a large transcriptomic screen of differentially vulnerable motor pools in SMA mice throughout disease progression in Paper II. We used laser capture microdissection (LCM) to dissect motor neurons from different pools in the brainstem and spinal cord. Here we revealed pathways and transcripts that were enriched only in the resistant oculomotor neurons that serve to prevent them from going into apoptosis, as well as promote their regeneration. We showed that applying one of the oculomotor-enriched transcripts, Gdf15, to vulnerable spinal motor neurons derived from stem cells improved their survival. Next, in Paper III we utilised in vitro approaches and generated a model of oculomotor resistance in ALS by deriving both spinal and oculomotor neurons from mouse embryonic stem cells. Here we found that in vitro oculomotor neurons are more resistant to excitotoxic stress and have increased levels of prosurvival Akt-signaling, which also held up in LCM-dissected post-mortem human spinal and oculomotor neurons.
To investigate the vulnerability of the distal axon, we generated a technique called Axon-Seq in Paper IV, that allows transcriptomic profiling of isolated motor axons by culturing motor neurons in microfluidic chambers. We subsequently used this technique in Paper V to show that human stem cell-derived motor neurons carrying ALS-causative mutations in FUS and TDP-43 showed unique transcriptome dysregulation in somas and axons. This implies that different pathways of degeneration are at play between subcellular compartments, and provides novel targets for therapeutic intervention directed at different compartments of the motor neuron.
We set out to investigate the mechanisms that underlie the resistance of oculomotor neurons, as well as the differential vulnerability within motor neurons themselves. We first validated the resistance of oculomotor neurons in two mouse models of ALS and SMA in Paper I. Next, we performed a large transcriptomic screen of differentially vulnerable motor pools in SMA mice throughout disease progression in Paper II. We used laser capture microdissection (LCM) to dissect motor neurons from different pools in the brainstem and spinal cord. Here we revealed pathways and transcripts that were enriched only in the resistant oculomotor neurons that serve to prevent them from going into apoptosis, as well as promote their regeneration. We showed that applying one of the oculomotor-enriched transcripts, Gdf15, to vulnerable spinal motor neurons derived from stem cells improved their survival. Next, in Paper III we utilised in vitro approaches and generated a model of oculomotor resistance in ALS by deriving both spinal and oculomotor neurons from mouse embryonic stem cells. Here we found that in vitro oculomotor neurons are more resistant to excitotoxic stress and have increased levels of prosurvival Akt-signaling, which also held up in LCM-dissected post-mortem human spinal and oculomotor neurons.
To investigate the vulnerability of the distal axon, we generated a technique called Axon-Seq in Paper IV, that allows transcriptomic profiling of isolated motor axons by culturing motor neurons in microfluidic chambers. We subsequently used this technique in Paper V to show that human stem cell-derived motor neurons carrying ALS-causative mutations in FUS and TDP-43 showed unique transcriptome dysregulation in somas and axons. This implies that different pathways of degeneration are at play between subcellular compartments, and provides novel targets for therapeutic intervention directed at different compartments of the motor neuron.
List of papers:
I. Comley LH, Nijssen J, Frost-Nylen J and Hedlund E. Cross-Disease Comparison of Amyotrophic Lateral Sclerosis and Spinal Muscular Atrophy Reveals Conservation of Selective Vulnerability but Differential Neuromuscular Junction Pathology. J Comp Neurol. 524(7):1424-1442
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II. Nichterwitz S, Nijssen J*, Storvall H*, Schweingruber C, Comley LH, Allodi I, van der Lee M, Deng Q, Sandberg R and Hedlund E. LCM-seq reveals unique transcriptional adaption mechanisms of resistant neurons and identifies protective pathways in spinal muscular atrophy. * These authors contributed equally to this work [Accepted]
Fulltext (DOI)
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III. Allodi I, Nijssen J, Aguila Benitez J, Schweingruber C, Fuchs A, Bonvicini G, Cao M, Kiehn O and Hedlund E. Modeling Motor Neuron Resilience in ALS Using Stem Cells. Stem Cell Rep. 12(6):1329-1341
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IV. Nijssen J*, Aguila J*, Hoogstraaten R, Kee N and Hedlund E. Axon-Seq Decodes the Motor Axon Transcriptome and Its Modulation in Response to ALS. Stem Cell Rep. 11(6):1565-1578 * These authors contributed equally to this work
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V. Nijssen J, Aguila J, Schweingruber C, Reber S, Ruepp MD and Hedlund E. Mutations in the ALS-causative genes FUS and TDP-43 cause distinct dysregulation of somatic and axonal transcriptomes. [Manuscript]
I. Comley LH, Nijssen J, Frost-Nylen J and Hedlund E. Cross-Disease Comparison of Amyotrophic Lateral Sclerosis and Spinal Muscular Atrophy Reveals Conservation of Selective Vulnerability but Differential Neuromuscular Junction Pathology. J Comp Neurol. 524(7):1424-1442
Fulltext (DOI)
Pubmed
View record in Web of Science®
II. Nichterwitz S, Nijssen J*, Storvall H*, Schweingruber C, Comley LH, Allodi I, van der Lee M, Deng Q, Sandberg R and Hedlund E. LCM-seq reveals unique transcriptional adaption mechanisms of resistant neurons and identifies protective pathways in spinal muscular atrophy. * These authors contributed equally to this work [Accepted]
Fulltext (DOI)
Pubmed
III. Allodi I, Nijssen J, Aguila Benitez J, Schweingruber C, Fuchs A, Bonvicini G, Cao M, Kiehn O and Hedlund E. Modeling Motor Neuron Resilience in ALS Using Stem Cells. Stem Cell Rep. 12(6):1329-1341
Fulltext (DOI)
Pubmed
View record in Web of Science®
IV. Nijssen J*, Aguila J*, Hoogstraaten R, Kee N and Hedlund E. Axon-Seq Decodes the Motor Axon Transcriptome and Its Modulation in Response to ALS. Stem Cell Rep. 11(6):1565-1578 * These authors contributed equally to this work
Fulltext (DOI)
Pubmed
View record in Web of Science®
V. Nijssen J, Aguila J, Schweingruber C, Reber S, Ruepp MD and Hedlund E. Mutations in the ALS-causative genes FUS and TDP-43 cause distinct dysregulation of somatic and axonal transcriptomes. [Manuscript]
Institution: Karolinska Institutet
Supervisor: Hedlund, Eva
Co-supervisor: Kee, Nigel
Issue date: 2020-05-06
Rights:
Publication year: 2020
ISBN: 978-91-7831-841-4
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