Epigenetic regulation of axonal regeneration

Authors: S. DI GIOVANNI, E. MACLACHLAN, I. PALMISANO, T. HUTSON, A. HERVERA, F. DE VIRGILIIS, M. DANZI, J. BIXBY, V. LEMMON

Professor Simone Di Giovanni, Imperial College, UK
Di Giovanni Imperial College of London

Lab Abstract:
Regeneration after peripheral nerve injury depends on the activation of key signalling events, the recruitment of transcription factors and histone acetylation. This does not occur after a central spinal injury, which is associated with regenerative failure. Whether specific histone marks and the recruitment of transcription factors confers a differential molecular signature between a central versus a peripheral axonal injury remained to be determined. Here, we performed RNAseq and ChIPseq for H3K27ac and H3K9ac in the sciatic dorsal root ganglia after central spinal cord or peripheral sciatic nerve injury, to identify key molecular events associated with regeneration vs regenerative failure. Data analysis showed that increased promoter enrichment of H3K27ac/H3K9ac occurs across various regenerative associated genes after sciatic injury only. Bioinformatics analysis identified a gene network enriched in transcription factors and histone acetylation.

Indeed, we found that this complex is required for the regenerative growth of sensory neurons including after a spinal injury. Lastly, we identified the FDA approved HDAC inhibitor Panobinostat that activates this network to promote axonal regeneration after spinal cord injury.

Abstract Citation
*S. DI GIOVANNI1, E. MACLACHLAN1, I. PALMISANO1, T. HUTSON1, A. HERVERA1, F. DE VIRGILIIS1,2, M. DANZI2, J. BIXBY2, V. LEMMON2;
1Imperial Col. London, London,, United Kingdom; 2University of Miami, Miami, FL. Epigenetic regulation of axonal regeneration. Program No. 213.15. 2018 Neuroscience Meeting Planner. San Diego, CA: Society for Neuroscience, 2018. Online.

Advances and Limitations of Current Epigenetic Studies Investigating Mammalian Axonal Regeneration Open Access in Neurotherapeutics Palmisano, I. & Di Giovanni, S. Neurotherapeutics (2018) 15: 529. https://doi.org/10.1007/s13311-018-0636-1

Posted in Chronic Spinal Cord Injury Research, Gene Therapy, Neuroscience Abstracts, Regenerative Medicine, spinal cord injury research, Stem Cell Research | Tagged , | Leave a comment

Tune into SCI 2020 tomorrow and Wednesday

The goal of the ‘SCI 2020: Launching a Decade for Disruption in Spinal Cord Injury Research’ conference is to initiate discussion across the SCI research community to launch a new decade of research that disrupts traditional barriers and brings about collaborative efforts to address the key research questions in spinal cord injury research. This conference is designed to be a comprehensive stakeholder’s meeting to bring diverse experience and voices together with this common goal. The participants will be challenged to critically evaluate the state of the science, assess areas of scientific, technological and community readiness, and identify the collaborations needed to change the trajectory of research and clinical opportunities for people with SCI.

Objectives:

To address and raise awareness of recent progress and current gaps in SCI research
To provide opportunities for collaboration across basic, translational, clinical research and consumer groups
To identify the top SCI research priorities for next 5-10 years — of and for the SCI research community — at the intersection of Scientific / Technological / Community Readiness

Live streaming and archived video will be available at: https://videocast.nih.gov/

The meeting will be open and available on videocast for those unable to attend in person on the NIH Campus.

Posted in Chronic Spinal Cord Injury Research, spinal cord injury research | Leave a comment

Engineered Neuroplasticity for Spinal Cord Rehabilitation

“Engineered Neuroplasticity for Spinal Cord Rehabilitation” by Chet Moritz, PhD, PT (University of Washington), at the 7th Annual International Symposium on Regenerative Rehabilitation.

 

Posted in Chronic Spinal Cord Injury Research, Neuromodulation, Regenerative Medicine, Rehabilitation, spinal cord injury research | Tagged , | Leave a comment

A novel biomaterial-based Epac targeting approach to promote spinal cord repair

Authors: A. GUIJARRO-BELMAR, M. VISKONTAS, X. BO, D. SHEWAN, W. HUANG

Alba Guijarro Belmar, PhD student at University of Aberdeen

Lab Abstract:
Spinal cord injury (SCI) is a highly debilitating trauma affecting millions of patients worldwide with no cure. The challenges for spinal repair include a lack of intrinsic capacity for adult CNS neurons to regrow post injury and the inhibitory physical and chemical barriers formed at the lesion site. One of the most promising new avenues for spinal repair is to combine novel axon growth-promoting and tissue-engineering strategies, thus creating a permissive and encouraging environment for injured spinal nerve processes to regrow.
Elevated levels of cAMP have been shown to promote injured CNS neurons to sprout and extend neurites, even in the presence of growth inhibitory molecules including chondroitin sulphate proteoglycans (CSPGs). cAMP is now known to signal via Epac, a guanine nucleotide exchange factor, which has two isoforms, Epac1 and Epac2, with the latter expressed mainly in postnatal neural tissue. By using primary neuronal cultures, immunocytochemistry and lentiviral tools, we herein demonstrate that specific activation of Epac2 significantly enhances neurite outgrowth of cultured postnatal rat cortical neurons and is able to overcome the inhibitory effects of CSPGs and mature activated astrocytes on cortical neuron growth in vitro.
With the aim to find an effective combinatorial strategy, we have further explored the suitability of a novel self-assembling hydrogel for spinal repair. The gel is a synthetic polymer with 3-D nanostructured networks that are similar to native extracellular matrix and can be functionalised with growth-promoting drugs. We show that this gel has appropriate stiffness, biocompatibility and biodegradation properties to promote cultured postnatal cortical neuron growth. By using an ex vivo model that mimics the in vivo environment after spinal injury, we demonstrate that the gel incorporated with a specific Epac2 agonist promotes significantly more axon regrowth than gel alone and without gel. By using a partial transection model of spinal injury, we further demonstrate that the spinal injured rats receiving implantation of the gel incorporated with the Epac2 agonist have much better locomotor function recovery than those rats having gel only implantation.
Our results demonstrate the potential of combining Epac2 activation with novel 3-D growth-supporting hydrogels to promote axon regrowth, thus contributing to new methods for spinal repair. Future studies will examine the efficacy of this combined strategy in a clinically relevant contusion model of spinal injury.

Grant Support: Nathalie Rose Barr Studentship. International Spinal Research Trust

Abstract Citation
*A. GUIJARRO-BELMAR1, M. VISKONTAS1, X. BO2, D. SHEWAN1, W. HUANG1;
1Univ. of Aberdeen, Aberdeen, United Kingdom; 2Ctr. for Neurosci. and Trauma, Queen Mary Univ. of London, London, United Kingdom. A novel biomaterial-based Epac targeting approach to promote spinal cord repair. Program No. 213.04. 2018 Neuroscience Meeting Planner. San Diego, CA: Society for Neuroscience, 2018. Online.

Posted in Biomaterials, Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Regenerative Medicine, spinal cord injury research | Tagged , | 5 Comments

Transplanting Cells for Spinal Cord Repair: Who, What, When, Where and Why?

Authors: L.V. Zholudeva, Michael.A. Lane

Lab Abstract:

“Cellular transplantation for repair of the injured spinal cord has a rich history with strategies focused on neuroprotection, immunomodulation, and neural reconstruction. The goal of the present review is to provide a concise overview and discussion of five key themes that have become important considerations for rebuilding functional neural networks. The questions raised include: (1) who are the donor cells selected for transplantation, (2) what is the intended target for repair, (3) when is the optimal time for transplantation, (4) where should the cells be delivered, and lastly (5) why does cell transplantation remain an attractive candidate for promoting neural repair after injury? Recent developments in neurobiology and engineering now enable us to start addressing these questions with multidisciplinary expertise and methods.”

See the Full Published Journal Article in Cell Transplantation: HERE

Zholudeva, L. V., & Lane, M. A. (2019). Transplanting Cells for Spinal Cord Repair: Who, What, When, Where and Why? Cell Transplantation.   https://doi.org/10.1177/0963689718824097

Lyandysha V. Zholudeva, BS Drexel University, Spinal Cord Injury Research Center

Funding:
The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was funded by the NINDS, NIH R01 NS081112, RO1 NS104291, The Moseley Foundation (Lane), Craig H. Neilsen (#338432, Lane), the Drexel Deans Fellowship for Collaborative or Themed Research (Zholudeva), and the Spinal Cord Research Center at Drexel University, College of Medicine.

Posted in Chronic Spinal Cord Injury Research, Regenerative Medicine, spinal cord injury research, Stem Cell Research | Tagged , | Leave a comment

Stem Cell Therapy for Traumatic SCI

In this presentation, Dr.Alicia Fuhrman in the Department of Rehabilitation Medicine, University of Washington, explains the complex and rapidly expanding field of stem cell medicine. She begins by clarifying what stem cells are and how they are used in different treatments. She then summarizes what the research can tell us about how effective these treatments are for spinal cord injuries. Finally, she discusses how to recognize and avoid scams and how to decide about treatment for yourself or, if you are a health provider, how to counsel your patients. After watching, please complete our 2-minute survey at https://is.gd/scivideos. Visit our website at http://sci.washington.edu.

 

Posted in Chronic Spinal Cord Injury Research, Regenerative Medicine, spinal cord injury research, Stem Cell Research | Tagged , | Leave a comment

Translational Regenerative Therapies for Chronic Spinal Cord Injury

Authors: Kyriakos Dalamagkas, Magdalini Tsintou, Amelia Seifalian, Alexander M. Seifalian

Abstract: “Spinal cord injury is a chronic and debilitating neurological condition that is currently being managed symptomatically with no real therapeutic strategies available. Even though there is no consensus on the best time to start interventions, the chronic phase is definitely the most stable target in order to determine whether a therapy can effectively restore neurological function. The advancements of nanoscience and stem cell technology, combined with the powerful, novel neuroimaging modalities that have arisen can now accelerate the path of promising novel therapeutic strategies from bench to bedside. Several types of stem cells have reached up to clinical trials phase II, including adult neural stem cells, human spinal cord stem cells, olfactory ensheathing cells, autologous Schwann cells, umbilical cord blood-derived mononuclear cells, adult mesenchymal cells, and autologous bone-marrow-derived stem cells. There also have been combinations of different molecular therapies; these have been either alone or combined with supportive scaffolds with nanostructures to facilitate favorable cell–material interactions. The results already show promise but it will take some coordinated actions in order to develop a proper step-by-step approach to solve impactful problems with neural repair.”

LINK: See the Full Open Access Review in the International Journal of Molecular Sciences 

Author Contributions: A.M.S. proposed the review article and M.T., K.D., and A.S. worked on data gathering and drafting the review. All four authors critically analyzed and reviewed the topic and completed the review with future direction of the research.

Acknowledgments: M.T. was supported by Onassis Foundation during that study.

Posted in Chronic Spinal Cord Injury Research, Regenerative Medicine, spinal cord injury research, Stem Cell Research | Tagged , , , | 3 Comments

Biomimetic 3D-printed scaffolds for spinal cord injury repair

Authors: Jacob Koffler, Wei Zhu, Xin Qu, Oleksandr Platoshyn, Jennifer N. Dulin, John Brock, Lori Graham, Paul Lu, Jeff Sakamoto, Martin Marsala, Shaochen Chen, Mark H. Tuszynski
“Current methods for bioprinting functional tissue lack appropriate biofabrication techniques to build complex 3D micrarchitectures essential for guiding cell growth and promoting tissue maturation . 3D printing of central nervous system (CNS) structures has not been accomplished, possibly owing to the complexity of CNS architecture. Here, we report the use of a microscale continuous projection printing method (μCPP) to create a complex CNS structure for regenerativmedicine applications in the spinal cord. μCPP can print 3D biomimetic hydrogel scaffolds tailored to the dimensions of the rodent spinal cord in 1.6 s and is scalable to human spinal cord sizes and lesion geometries. We tested the ability of µCPP 3D-printed scaffolds loaded with neural progenitor cells (NPCs) to support axon regeneration and form new ‘neural relays’ across sites of complete spinal cord injury in vivo in rodents. We find that injured host axons regenerate into 3D biomimetic scaffolds and synapse onto NPCs implanted intthe device and that implanted NPCs in turn extend axons out of the scaffold and into the host spinal cord below the injury to restore synaptic transmission and significantly improve functional outcomes. Thus, 3D biomimetic scaffolds offer a means of enhancing CNS regeneration through precision medicine.”

See the full open access online publication in Nature Medicine.

Posted in Biomaterials, Chronic Spinal Cord Injury Research, Regenerative Medicine, spinal cord injury research, Stem Cell Research | Tagged , , , , , , , , , , , | 2 Comments

Calcium imaging of synaptic connectivity between host and neural progenitor cell graft-derived neurons after spinal cord injury

Authors:
STEVEN L. CETO, K. SEKIGUCHI, A. NIMMERJAHN, M. H. TUSZYNSKI;

Steven Ceto,
Predoctoral Graduate Student

Lab Abstract:
Neural stem cells (NSCs) grafted into sites of spinal cord injury (SCI) may act as new electrophysiological relays between host neurons above and below the lesion. Host axons regenerate robustly into NSC grafts and form synapses; in turn, graft axons extend long distances into host white and gray matter caudal to the injury and form synapses. To investigate potential functionality of these new synaptic pathways, we performed calcium imaging and whole-cell patch clamp recordings in mice with NSC grafts after SCI. We placed T12 dorsal column lesions and acutely grafted embryonic day twelve (E12)-derived spinal cord neural progenitor cells (NPCs) expressing the calcium indicator GCaMP6f into the lesion site. From 6 to 8 weeks later, we imaged the activity of populations of neurons within NPC grafts in acute spinal cord slices, anesthetized, or awake behaving animals. After grafting NPCs into acute spinal cord injuries in mice, we imaged the simultaneous activity of populations of neurons within grafts both in vivo and in ex vivo slice preparations. We observed spontaneous activity in both neurons and glia, as well as hindpaw pinch- and cold air puff-evoked responses. Furthermore, optogenetic stimulation of corticospinal tract axons regenerating into grafts evoked robust responses throughout grafts. Activity patterns included large-scale events and independent, single-neuron activity. Spontaneous activity and stimulus-evoked responses were significantly enhanced by bath application of the potassium channel blocker 4-aminopyridine (4-AP). We are currently optimizing methods to interrogate graft-graft and graft-to-host connectivity. These studies will reveal the extent to which, at the cellular level, current NSC grafts are capable of forming functional neuronal relays across spinal cord injuries.

Grant Support
Wings for Life Individual Research Grant
Grant Support
UCSD Frontiers of Innovation Scholars Program
Grant Support
The Veterans Administration
Grant Support
Nakajima Foundation

Abstract Citation
*S. L. CETO1, K. SEKIGUCHI2, A. NIMMERJAHN3, M. H. TUSZYNSKI4;
1UCSD, LA Jolla, CA; 2Waitt Advanced Biophotonics Ctr., Salk Inst. for Biol. Sci., La Jolla, CA; 3Salk Inst. For Biol. Studies, La Jolla, CA; 4Univ. of California San Diego Dept. of Neurosciences, La Jolla, CA. Calcium imaging of synaptic connectivity between host and neural progenitor cell graft-derived neurons after spinal cord injury. Program No. 138.12. 2018 Neuroscience Meeting Planner. San Diego, CA: Society for Neuroscience, 2018. Online.

Posted in Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Regenerative Medicine, spinal cord injury research, Stem Cell Research | Tagged ,

Regaining trunk stability after spinal cord injury

Authors: M. RATH, D. G. SAYENKO, Y. P. GERASIMENKO, V. EDGERTON

Mrinal ‘Neil’ Rath, Graduate Student, UCLA.

Lab Abstract:
Recently we have developed the non-invasive electrical spinal stimulation technology for postural control in SCI subjects during standing. However, the potential of non-invasive spinal stimulation to facilitate trunk postural control during sitting in humans with spinal cord injury (SCI) has not been investigated. We hypothesized that transcutaneous electrical stimulation of the lumbosacral enlargement can improve trunk posture. Six participants with non-progressive SCI, C3-T9, AIS A or C, performed different motor tasks during sitting on a force platform. Electromyography of the trunk muscles, three-dimensional kinematics, and force plate data were acquired. Spinal stimulation improved trunk control during sitting in all tested individuals. Stimulation resulted in elevated activity of the erector spinae, rectus abdominis, and external obliques, contributing to trunk control, more natural anterior pelvic tilt and lordotic curve, and greater multidirectional seated stability. During spinal stimulation prior to any training, the center of pressure (COP) excursion decreased to 112.06 ± 36.00 mm from 143.74 ± 30.79 mm (p=.028, Z=-2.2014) without stimulation and to 93.09 ± 37.42 mm from 123.77 ± 48.44 mm (p=.028, Z=-2.2014) without stimulation in quiet sitting before training and after training respectively. Similarly, the limits of stable displacement increased by 31.40 ± 37.28% (p=.046, Z=1.9917), 19.42 ± 15.83% (p=.046, Z=1.9917), 54.11 ± 54.36% (p=.028, Z=2.2014), and 49.69 ± 32.343% (p=.046, Z=1.9917) before training and 24.06 ± 16.06% (p=.028, Z=2.2014), 20.25 ± 22.01% (p=.075, Z= 1.7821), 27.87 ± 11.88% (p=.028, Z=2.2014), and 27.33 ± 44.42% (p=.116, Z= 1.5724) after training in the forward, backward, right, and left directions, respectively. These data demonstrate that the spinal networks can be modulated transcutaneously with tonic electrical spinal stimulation to physiological states sufficient to generate a more stable, erect sitting posture after chronic paralysis.

Grant Support: Paralyzed Veterans of America (PVA) Research Foundation Grant #3068, NIH SBIR Grant R43EB018232, Russian Foundation for Fundamental Research Grant 16-29-08173-ofi-m

Disclosures:
M. Rath: None. D.G. Sayenko: None. Y.P. Gerasimenko: E. Ownership Interest (stock, stock options, royalty, receipt of intellectual property rights/patent holder, excluding diversified mutual funds); shareholder interest in NeuroRecovery Technologies. V. Edgerton: E. Ownership Interest (stock, stock options, royalty, receipt of intellectual property rights/patent holder, excluding diversified mutual funds); shareholder interest in NeuroRecovery Technologies.

Abstract Citation
*M. RATH1, D. G. SAYENKO1, Y. P. GERASIMENKO2, V. EDGERTON3;
1UCLA, Los Angeles, CA; 2Pavlov Inst. of Physiol, St Petersburg, Russian Federation; 3Dept Integrative Biol. & Physiol., Univ. of California Los Angeles, Los Angeles, CA. Regaining trunk stability after spinal cord injury. Program No. 138.16. 2018 Neuroscience Meeting Planner. San Diego, CA: Society for Neuroscience, 2018. Online.

Posted in Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Rehabilitation, spinal cord injury research | Tagged , | 6 Comments