Interaction of reactive astrocytes with type I collagen induces astrocytic scar formation through the integrin–N-cadherin pathway after SCI

Masamitsu Hara Department of Advanced Medical Initiatives, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.

Central nervous system (CNS) injury transforms naive astrocytes into reactive astrocytes, which eventually become scar-forming astrocytes that can impair axonal regeneration and functional recovery. This sequential phenotypic change, known as reactive astrogliosis, has long been considered unidirectional and irreversible. However, we report here that reactive astrocytes isolated from injured spinal cord reverted in retrograde to naive astrocytes when transplanted into a naive spinal cord, whereas they formed astrocytic scars when transplanted into injured spinal cord, indicating the environment-dependent plasticity of reactive astrogliosis. We also found that type I collagen was highly expressed in the spinal cord during the scar-forming phase and induced astrocytic scar formation via the integrin–N-cadherin pathway. In a mouse model of spinal cord injury, pharmacological blockade of reactive astrocyte–type I collagen interaction prevented astrocytic scar formation, thereby leading to improved axonal regrowth and better functional outcomes. Our findings reveal environmental cues regulating astrocytic fate decisions, thereby providing a potential therapeutic target for CNS injury.

Masamitsu Hara1,2, Kazu Kobayakawa2 , Yasuyuki Ohkawa3 , Hiromi Kumamaru2, Kazuya Yokota2, Takeyuki Saito1,2, Ken Kijima1,2, Shingo Yoshizaki1,2, Katsumi Harimaya2, Yasuharu Nakashima2 & Seiji Okada1,2

Nature Medicine (2017) doi:10.1038/nm.4354
Received 29 September 2016 Accepted 14 April 2017 Published online 19 June 2017

Posted in Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Spinal Research, Stem Cell Research | Tagged ,

SCI Stem-cell pioneer enters the political field in California

Democrat Hans Keirstead is one of five Democrats challenging Rep. Dana Rohrabacher, R-Costa Mesa.

Hans Keirstead, a pioneering stem-cell biologist and multimillionaire businessman who was a professor at UC Irvine for 14 years, announced Thursday he would join the field of Democrats challenging Rep. Dana Rohrabacher, R-Costa Mesa.

Keirstead said the leading motivation for his decision to run was the country’s health care system, with specific concerns about Republican plans that could reduce those with insurance by 23 million those with insurance and with regulatory fee hikes for companies doing lifesaving research.

“That is going to have a direct effect on development in the biomedical field,” the Laguna Beach resident said. “There’s an ignorance in Washington about health care … an ignorance of the system.”

Keirstead, 50, is a Canadian immigrant who sold his Irvine-based California Stem Cell Inc. for more than $100 million in 2014. His biography at Aivita Biomedical, where he’s currently CEO, says that he raised $93 million in grants and donations while at UC Irvine and $22 million in grants at his current job.

Orange County Register: Martin Wisckol LINK

Posted in Advocacy | Tagged

Patients with Complete Paralysis Show Additional Recovery of Arm, Hand and Finger Function at 9-months After Treatment with Asterias’ AST-OPC1

Third patient recovers two motor levels; three of six (50%) patients in AIS-A 10 million cell cohort have now recovered two motor levels on at least one side

Asterias OPC.


FREMONT, Calif., June 13, 2017 /PRNewswire/ –Asterias Biotherapeutics, Inc. (NYSE MKT: AST), a biotechnology company pioneering the field of regenerative medicine, today announced that new 9-month follow-up data from the AIS-A 10 million cell cohort in the company’s ongoing SCiStar Phase 1/2a clinical trial shows three of six (50%) patients have now recovered two levels of motor function and previously-announced improvements in arm, hand and finger function at 3-months and 6-months following administration of AST-OPC1 have been confirmed and further increased at 9-months.

“The new efficacy results show that previously reported meaningful improvements in arm, hand and finger function in the 10 million cell cohort treated with AST-OPC1 cells have been maintained and in some patients have been further enhanced even 9 months following dosing,” stated Dr. Edward Wirth III, Chief Medical Officer. “Gains in motor function, such as the improvements observed in the SCiStar study to date, have been shown to increase a patient’s ability to function independently following complete cervical spinal cord injuries. We are increasingly encouraged by these continued positive results, which are remarkable compared with spontaneous recovery rates observed in a closely matched untreated patient population.”

Jane S. Lebkowski, Ph.D., Asterias’ President of R&D and Chief Scientific Officer, will present the 9-month efficacy and safety data on the AIS-A 10 million cell cohort later today during the International Society for Stem Cell Research (ISSCR) 2017 Annual Meeting held in Boston, MA. The full slide presentation will be available at http://asteriasbiotherapeutics.com/inv_events_presentations.php.

Posted in Chronic Spinal Cord Injury Research, Regenerative Medicine, Spinal Research, Stem Cell Research | Tagged , , | 2 Comments

Induction of immune tolerance by short-course immunosuppresion after spinal grafting of allogeneic neural precursors in pigs with previous chronic spinal cord traumatic injury.

Current clinical protocols use transient or continuous immunosuppression in patients receiving allogeneic neural precursor grafts for treatment of a variety of neurological diseases, including spinal trauma, stroke or ALS. At present there is no solid evidence, however, that would confirm long-term immune tolerance to allogeneic grafts in a large animal model(s) of chronic spinal cord injury. In our current study, we have tested the engraftment of porcine fetal NPCs after transplantation into the lumbar spinal cord of allogeneic transiently-immunosuppressed spinally-injured recipients. Material and Methods: Porcine fetal spinal cord-derived NPCs were isolated from 30-day-old fetuses. NPCs were expanded and characterized by immunofluorescence staining after differentiation in vitro. Clones of UBI-GFP expressing NPCs were then prepared and used in vivo for grafting into the lumbar spinal cord of allogeneic pigs with previous spinal cord L3 contusion injury. Animals (n=4) with fully developed paraplegia were grafted at 5.5 months after spinal injury. All animals received between 30-40 injections of NPCs targeted above, at epicenter and below the injury (10 ul/300,000 cells/injection). After cell grafting, animals were immunosuppressed iv with a combination of Prograf and MFF for 4 weeks. At 4 weeks, immunosuppression was terminated and animals survived for an additional 3 months (n=1), 6 months (n=1) or 12 months (n=2) without immunosuppression. After survival, animals were perfusion fixed and the presence and differentiation of grafted cells analyzed by immunofluorescence. Results: i) In vitro induced NPCs showed expression of neural/neuronal markers, including DCX, NeuN, GABA and GFAP. ii) Analysis of previously injured spinal cord tissue grafted with UBI-GFP+ NPCs showed extensive GFP+ grafts occupying previously injured spinal cord regions. iii) Double staining with neuronal markers (NeuN, SYN, NSE) showed the presence of high density grafted neurons throughout the grafts. iv) Staining with excitatory and inhibitory neurotransmitter markers showed a preferential GABA/glycin-ergic phenotype (VGAT and GAD65 +) in grafted neurons. v) Staining with glial markers showed near complete repopulation of individual grafts with GFAP+ astrocytes and regularly distributed Olig2 positive oligodendrocytes at 12 months after grafting. vi) No signs of cell-mediated rejection were seen at any time point after cell grafting. Conclusion: These data demonstrate that short course (4 weeks) immunosuppression is effective in inducing immune tolerance to allogeneic NPCs after grafting into chronically-injured spinal cord in pigs.

Abstract Authors
*M. MARSALA1, J. D. CIACCI2, E. I. CURTIS2, S. MARSALA1, M. R. NAVARRO1, P. CHEN1, S. JUHAS3, J. JUHASOVA3, K. YAMADA4, K. JOHE5;
1Dept. of Anesthesiol., 2Dept. of Neurosurg., Univ. of California San Diego, La Jolla, CA; 3Lab. of Cell Regeneration and Plasticity, Inst. of Animal Physiol. and Genet., Libechov, Czech Republic; 4Columbia Univ. Med. Ctr., New York, NY; 5Neuralstem, Germantown, MD
Disclosures
M. Marsala: None. J.D. Ciacci: None. E.I. Curtis: None. S. Marsala: None. M.R. Navarro: None. P. Chen: None. S. Juhas: None. J. Juhasova: None. K. Yamada: None. K. Johe: None.

LINK: Session 320 – Injury Responses after Spinal Cord Injury

Posted in Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Regenerative Medicine, Rehabilitation, Spinal Research, Stem Cell Research | Tagged | 1 Comment

Growth Hormone Improves Sensory Function in Complete Spinal Injury

Deficiency Growth Hormone and Spinal Cord Injury: In March 2013, the Spanish Medicines Agency approved a pilot clinical trial to investigate the efficacy of growth hormone (GH) as an adjunct to intensive rehabilitation in patients with spinal cord injury and GH deficiency (SCI-GHD-201 study).

By Becky McCall May 24, 2017

Dr. Guillem Cuatrecasas
Endocrinology
CPEN Coordinator

LISBON, Portugal — Six months of treatment with growth hormone has improved sensory function in patients with spinal-cord injury and concomitant growth-hormone deficiency, according to the first such trial of this approach.
“Changes in sensory quantification (electrical perception threshold) of up to five levels below the site of spinal injury were observed,” reported Gulliem Cuatrecasas, MD, PhD, an endocrinologist from Hospital Quiron-Teknon, Barcelona, Spain, who presented the work here at the European Congress of Endocrinology (ECE) 2017.

“This study looked at complete lesions, which are the severest form of spinal lesion, so we didn’t really expect to find any changes,” he stressed. Although these findings seem quite remarkable, it’s important that they are interpreted cautiously, “because it is not a solution for spinal-injury lesions. These patients do not walk again,” he stressed. But they may significantly affect quality of life. With these changes of up to five levels of improvement in sensation, “they may feel [the fact] that they are in a wheelchair, or they may feel a burning sensation or similar,” which may, for example, help to avoid pressure ulcers from wheelchair use.

And “autonomic nervous system processes also improved with less need for catheterization of the bladder after treatment with growth hormone,” Dr Cuatrecasas noted.

See the full article at Medscape

Posted in Chronic Spinal Cord Injury Research, Regenerative Medicine, Rehabilitation, Spinal Research | Tagged

Promoting targeted reinnervation of phrenic motor neurons and restoration of respiratory function using BDNF after SCI

Brittany Charsar:
Laboratory of Angelo Lepore, PhD


We are working a novel approach to promote regrowth of damaged descending bulbospinal respiratory axons and reinnervation of their correct phrenic motor neuron (PhMN) targets after cervical spinal cord injury (SCI). Cervical SCI, which occurs in more than half of all human cases, can be extremely debilitating if the neural circuitry responsible for controlling respiratory function is affected. PhMNs located at cervical levels C3-C5 directly control activation of the diaphragm, which is the major inspiratory muscle. PhMNs are mono-synaptically innervated by bulbospinal projections of respiratory neurons located in a brainstem nucleus called the rostral Ventral Respiratory Group (rVRG). Cervical SCI can result in persistent diaphragm compromise because of damage to these descending rVRG axons, denervation and silencing of spared PhMNs, and consequent paralysis of the hemi-diaphragm. In a rat model of unilateral C2/3 hemisection SCI, we are expressing the axon guidance molecule, brain-derived neurotrophic factor (BDNF), in PhMNs to direct regenerating ipsilateral and/or sprouting contralateral rVRG axons towards PhMNs with the goal of achieving targeted restoration of the critical rVRG-PhMN-diaphragm circuit. Specifically, we are employing anatomically-targeted delivery of adeno-associated virus serotype 2 (AAV2) to the ipsilateral C3-C5 spinal cord to achieve BDNF expression throughout the denervated PhMN pool. Using neuroanatomical tract tracing and in vivo electrophysiological approaches, we are exploring the effects of this strategy on rVRG axon regrowth and collateral sprouting, synaptic reconnection with PhMNs, and restoration of ipsilateral hemi-diaphragm activity. Given our findings that rVRG neurons express the BDNF receptor, tropomyosin-related kinase B (TrkB), and that we can efficiently transduce PhMNs using this AAV2-based approach, we hypothesize that BDNF will promote robust PhMN reinnervation by injured rRVG axons and diaphragmatic respiratory recovery.

Abstract Authors:
*B. CHARSAR1, M. URBAN1, B. GHOSH1, G. M. SMITH2, A. C. LEPORE1;
1Thomas Jefferson Univ., Philadelphia, PA; 2Temple Univ., Philadelphia, PA
Disclosures
B. Charsar: None. M. Urban: None. B. Ghosh: None. G.M. Smith: None. A.C. Lepore: None.

LINK: Session 320 – Injury Responses after Spinal Cord Injury

RESPIRATORY MOTOR NEURON PROTECTION FOLLOWING CERVICAL SPINAL CORD INJURY (NIH RePorter LINK)

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

New Pathways for Recovery of Function Following Paralysis

We are dedicated to developing disease and injury specific devices to help restore optimal physiology function in individuals suffering with paralysis due to injury and illness. We accomplish this by enabling the reactivation of damaged and dormant neural circuits through neuromodulation of the spinal cord. The method we use is to apply our proprietary stimulator technology below the level of injury targeting neural connections & pathways with our discovered stimulation parameters enabling the spinal cord to re-awaken and recall what it is suppose to do. NeuroRecovery Technologies Inc.

LINK: Feed-Forwardness of Spinal Networks in Posture and Locomotion

LINK: Electrical neuromodulation of the cervical spinal cord facilitates forelimb skilled function recovery in spinal cord injured rats.

 

 

Posted in Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Rehabilitation, Spinal Research | Tagged , | 1 Comment

The Impact of combination biologics on treadmill quadrupedal locomotion after spinal cord hemi-contusion in non-human primates

Introduction.
Non-human primates can be trained to ambulate on a treadmill. Cervical contusions lead to both upper and lower extremity functional deficits. These can be analyzed in a coupled manner using kinematic tracking of forelimb and hindlimb joint cycles. We tested the following hypotheses. 1) Kinematic analysis has sufficient sensitivity to detect differences in injury severity after C3/4 hemi-contusion. 2) The impact of locally delivered experimental therapeutics including autologous Schwann cells (aSC) and lentivirus expressing chondroitinase ABC (LV-ChABC) on gait is evident on limb-coupled kinematic analysis. 3) This analysis adds relevant data not evident in hand dexterity testing.
Methods.
Five monkeys (M. fascicularis) were pre-trained. Following a right sided hemi-contusion the animals were randomized into A) No treatment controls (n= 1); B) LV-ChABC injected perilesionally 2 hours post spinal cord injury (n=2); C) LV-ChABC injection and transplant of aSC 14 days post-injury (n=2). Treadmill activity occurred weekly. Quadrupedal locomotion analysis was performed pre-injury, 3 and 6 months post-injury. Joints in the left and right sides were marked with ultraviolet ink and data was captured using black lights and a Vicon Motus tracking system. The variables assessed were joint track consistency and distance, stride length, and height.
Results.
All animals, except those receiving LV-ChABC, had notable deficits in quadrupedal locomotion at 3 months post-injury (step height, length). The wrist and ankle joint cycles were inconsistent possibly due to impaired strength, proprioception, and balance. By six months post-injury the joint cycles of groups A and B were more consistent and approached baseline. Animals in group C (aSC) showed persistent gait impairments. Analysis of contusion parameters including force delivered and ultrasound quantitative assessment of injury volume do not account for the behavioral differences.
Conclusion.
Kinematic quadrupedal locomotor assessment is useful to quantify recovery, adding to assessments of hand dexterity. The animals continue to survive. Final MRI, histology, and CST tracing will be correlated to the quadrupedal kinematic analysis.

Authors
*R. DE NEGRI1, A. J. SANTAMARIA1, F. D. BENAVIDES1, A. Y. FLORES1, N. JAMES4, Y. NUNEZ2, J. P. SOLANO2, J. VERHAAGEN5,6, E. J. BRADBURY4, J. D. GUEST1,3;
1The Miami Project to Cure Paralysis, 2Pedriatic Critical Care, 3Neurolog. Surgery, Univ. of Miami, Miller Sch. of Med., Miami, FL; 4The Wolfson Ctr. for Age-Related Dis., King’s Col. London, London, United Kingdom; 5Dept. of Mol. and Cell. Neurobio., Vrije Univ. Amsterdam, Ctr. for Neurogenomics and Cognition research, Amsterdan, Netherlands; 6Lab. for Neuroregeneration, Netherlands Inst. for Neurosci., Amsterdam, Netherlands
Disclosures
R. De Negri: None. A.J. Santamaria: None. F.D. Benavides: None. A.Y. Flores: None. N. James: None. Y. Nunez: None. J.P. Solano: None. J. Verhaagen: None. E.J. Bradbury: None. J.D. Guest: None.

LINK: Session 158 – Spinal Cord Injury and Plasticity

Posted in Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Regenerative Medicine, Rehabilitation, Spinal Research | 1 Comment

Breakthrough Regenerative Therapeutics Company Establishes Scientific Advisory Board

Fortuna Fix Inc. (“Fortuna”), a private, clinical-stage biotech company, is aiming to be the first to eliminate the need for embryonic and fetal stem cells by using direct reprogramming of autologous cells to treat neurodegenerative diseases. Fortuna announced today the launch of its Scientific Advisory Board (“SAB”) with Professor Michael Fehlings, MD, PhD; Father Kevin FitzGerald, S.J., PhD; Col. (R) Dallas Hack, MD, MPH; and Professor James Giordano, PhD.

“We are excited and honored to have these world-leading experts join our SAB,” says CEO Jan-Eric Ahlfors. “We look forward to working with them to bring our novel regenerative medicine solutions to patients suffering from neurotrauma and neurodegeneration.”

Jan-Eric Ahlfors CEO and Chief Scientific Officer

Fortuna’s two flagship technologies — autologous directly reprogrammed neural precursor cells (“drNPC”) and Regeneration Matrix (“RMx™”) — are poised to lead a revolution in neuro-regeneration.

For the first time, patients suffering from neurotrauma or neurodegeneration will be able to get treated with autologous neural stem cells produced by direct reprogramming (i.e. starting with and only using the patient’s own cells, bypassing use of pluripotent stem cells and avoiding harvesting and use of human embryos or fetuses). The method of direct reprogramming developed by Fortuna relies on an ethical, rapid, high throughput, low cost and fully automated manufacturing process. As drNPC do not involve any genetic engineering, pluripotent stem cells, or use of immune-suppression, it provides patients with personalized stem cells that are also expected to have a greater safety profile. In addition, drNPC are expected to replace dead neural cells, something that no other current technology can do effectively.

RMx™ is a unique and highly efficient bio-scaffold for the promotion of neural tissue regrowth.

“Our testing of drNPC at the Krembil Neuroscience Centre of the University Health Network in various Spinal Cord Injury (“SCI”) animal models to characterize their regenerative capacity and safety profile indicates that drNPC are a promising source of therapeutic stem cells with potential for tissue preservation and functional improvement after SCI. I am highly encouraged by the reprogramming efficiency of drNPC and look forward to leading the clinical development of drNPC for SCI,” says Professor Fehlings, after working on the drNPC in his lab for two years.

Dr. Hack further remarks: “Fortuna’s autologous drNPC represent a major advance in cell therapy for treatment of CNS injury and degeneration. For the first time, neurons, astrocytes and oligodendrocytes — the three type of cells of the brain and spinal cord — can be repaired and replaced where these cells have died or been destroyed due to trauma or neurodegenerative disease. Fortuna’s proprietary automated manufacturing addresses a key hurdle of personalized cell therapy, making drNPC commercially viable both at small and large scale”

“Stem cell therapeutics have been plagued with controversy and hype, raising ethical and political issues that have resulted in a relatively hostile funding environment for research and development in the field. I am excited to work alongside Fortuna to help advance development of their ethical and commercially viable platform for cell therapeutics to benefit patients, their families, and our entire society,” says Father FitzGerald.

The SAB members encompass unique expertise in key areas of importance for the company:

About Fortuna Fix Inc.

Fortuna is a private, clinical-stage biotech company with a patented direct cell reprogramming technology platform together with a patented bio-scaffolding technology for treatment of neurodegenerative diseases and neurotrauma. The company is focused on clinical development of its platforms for a range of neurodegenerative diseases including SCI, Parkinson’s disease, stroke, TBI, and ALS. The company has developed a proprietary fully automated GMP manufacturing system for production of drNPC, initially to be used in clinical trials in Parkinson’s disease and Spinal Cord Injury.

Read the Full NewsWire Press Release: 

Fortuna Fix Website Link:

LINK: Working 2 Walk 2015 Presentations  Part 1.(Science Time): First-in man Clinical Trials on Directly Re-programmed Autologous Neural Stem Cells

LINK: Working 2 Walk 2015 Presentations Part 2. Jan-Eric Ahlfors Human Clinical Studies

Posted in Biomaterials, Chronic Spinal Cord Injury Research, Regenerative Medicine, Rehabilitation, Spinal Research, Stem Cell Research, Unite 2 Fight Paralysis | Tagged , , , , | 2 Comments

Oxygen improves blood flow, restores more function in spinal cord injuries

Karim Fouad and post-doctoral fellows Yaqing Li (center) and Ana M. Lucas-Osma (right) and their team made a new discovery that could alter how we view spinal cord function and rehabilitation after spinal cord injuries.
Credit: Laurie Wang, University of Alberta

A new discovery at the University of Alberta will fundamentally alter how we view spinal cord function and rehabilitation after spinal cord injuries. Neuroscientists found that spinal blood flow in rats was unexpectedly compromised long after a spinal cord injury (chronically ischemia), and that improving blood flow or simply inhaling more oxygen produces lasting improvements in cord oxygenation and motor functions, such as walking.

Previous work had shown that while blood flow was temporarily disrupted at the injury site, it resumed rapidly, and it was more or less assumed that the blood flow was normal below the injury. This turns out to be wrong.

The Edmonton Journal with David Bennett Video

See the Full News Article at Science Daily HERE

Journal Reference:
Yaqing Li, Ana M Lucas-Osma, Sophie Black, Mischa V Bandet, Marilee J Stephens, Romana Vavrek, Leo Sanelli, Keith K Fenrich, Antonio F Di Narzo, Stella Dracheva, Ian R Winship, Karim Fouad, David J Bennett. Pericytes impair capillary blood flow and motor function after chronic spinal cord injury. Nature Medicine, 2017; DOI: 10.1038/nm.4331

Karim Fouad Links

Posted in Chronic Spinal Cord Injury Research, Neuroscience Abstracts, Regenerative Medicine, Rehabilitation, Spinal Research | Tagged , , , , , , , , , , , ,