Minimally Invasive Injection to the Phrenic Nerve in a Porcine Hemidiaphragmatic Paralysis Model: A Pilot Study

Neurosurgery ◽  
2019 ◽  
Vol 87 (4) ◽  
pp. 847-853 ◽  
Author(s):  
Pavlos Texakalidis ◽  
Muhibullah S Tora ◽  
Skyler Canute ◽  
Nathan Hardcastle ◽  
Kelly Poth ◽  
...  
Keyword(s):  
Motor Neurons ◽  
Phrenic Nerve ◽  
Vital Signs ◽  
Swine Model ◽  
Large Animal ◽  
Diaphragmatic Dysfunction ◽  
Thoracoscopic Approach ◽  
Large Animal Models ◽  
Cord Injury ◽  
Motor Neuron Loss

Abstract BACKGROUND Neurodegenerative diseases and spinal cord injury can affect respiratory function often through motor neuron loss innervating the diaphragm. To reinnervate this muscle, new motor neurons could be transplanted into the phrenic nerve (PN), allowing them to extend axons to the diaphragm. These neurons could then be driven by an optogenetics approach to regulate breathing. This type of approach has already been demonstrated in the peripheral nerves of mice. However, there is no established thoracoscopic approach to PN injection. Also, there is currently a lack of preclinical large animal models of diaphragmatic dysfunction in order to evaluate the efficacy of potential treatments. OBJECTIVE To evaluate the feasibility of thoracoscopic drug delivery into the PN and to assess the viability of hemidiaphragmatic paralysis in a porcine model. METHODS Two Landrace farm pigs underwent a novel procedure for thoracoscopic PN injections, including 1 nonsurvival and 1 survival surgery. Nonsurvival surgery involved bilateral PN injections and ligation. Survival surgery included a right PN injection and transection proximal to the injection site to induce hemidiaphragmatic paralysis. RESULTS PN injections were successfully performed in both procedures. The animal that underwent survival surgery recovered postoperatively with an established right hemidiaphragmatic paralysis. Over the 5-d postoperative period, the animal displayed stable vital signs and oxygenation saturation on room air with voluntary breathing. CONCLUSION Thoracoscopic targeting of the porcine PN is a feasible approach to administer therapeutic agents. A swine model of hemidiaphragmatic paralysis induced by unilateral PN ligation or transection may be potentially used to study diaphragmatic reinnervation following delivery of therapeutics.

scholarly journals How to generate graded spinal cord injuries in swine?: Tools and procedures

2021 ◽  
Author(s):  
Mark Züchner ◽  
Manuel J. Escalona ◽  
Lena Hammerlund Teige ◽  
Evangelos Balafas ◽  
Lili Zhang ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Animal Models ◽  
Functional Outcome ◽  
Spinal Cord Injuries ◽  
Large Body ◽  
Swine Model ◽  
Large Animal ◽  
Large Animal Models ◽  
Cord Injury ◽  
The Impact

Spinal cord injury (SCI) is a medically, psychologically and socially disabling condition. A large body of our knowledge on the basic mechanisms of SCI has been gathered in rodents. For preclinical validation of promising therapies, the use of animal models that are closer to humans has several advantages. This has promoted a more intensive development of large animal models for SCI during the past decade. We have recently developed a multimodal SCI apparatus for large animals that generated biomechanically reproducible impacts in vivo. It is composed of a spring-load impactor and support systems for the spinal cord and the vertebral column. We now present the functional outcome of farm pigs and minipigs injured with different lesion strengths. There was a correlation between the biomechanical characteristics of the impact, the functional outcome, and the tissue damage observed several weeks after injury. We also provide a detailed description of the procedure to generate such a SCI in both farm pigs and minipigs, in the hope to ease the adoption of the swine model by other research groups.

The Hemisection Approach in Large Animal Models of Spinal Cord Injury: Overview of Methods and Applications

2018 ◽  
Vol 33 (3) ◽  
pp. 240-251
Author(s):  
S. Wilson ◽  
S. J. Nagel ◽  
L. A. Frizon ◽  
D. C. Fredericks ◽  
N. A. DeVries-Watson ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Animal Models ◽  
Large Animal ◽  
Large Animal Models ◽  
Cord Injury

scholarly journals TMOD-17. A SWINE MODEL OF GLIOBLASTOMA INDUCED BY SOMATIC GENE MODIFICATION

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi266-vi266
Author(s):  
Barbara Tschida ◽  
Dylan Duerre ◽  
Mandy Taisto ◽  
Staci Solin ◽  
Adrienne Watson
Keyword(s):  
Signaling Pathways ◽  
Gene Editing ◽  
Brain Size ◽  
Rapid Development ◽  
Swine Model ◽  
Large Animal ◽  
Induced Mutations ◽  
Gene Modification ◽  
Large Animal Models ◽  
Human Gbm

Abstract Glioblastoma (GBM) is the most common and malignant primary brain tumor. New, effective treatments for GBM are needed since the standard of care universally fails to cure patients. A major hurdle for GBM therapeutic development is the lack of relevant large animal models with high translational value for preclinical studies. To overcome this, we are developing a model of GBM in outbred, immune-proficient swine which have comparable brain size and anatomy to humans. GBM tumors will be driven by somatic alterations to major signaling pathways frequently altered in human GBM and will express a secreted reporter for tumor growth detectable in the peripheral blood. These alterations and reporter expression cassettes will be induced by delivering transposons and CRISPR/Cas9 gene-editing tools to the subventricular brain cells of live swine by stereotactic injections. We have developed and deployed these gene-editing tools, demonstrating their efficacy in swine cells. We have identified stereotactic coordinates to reproducibly target the lateral ventricles of neonatal swine brains and have demonstrated successful plasmid delivery to cells at these coordinates. Stable, persistent transposon integration and clonal expansion of modified cells has been confirmed by sequencing transposon-genomic DNA junctions in brain tissue six-months post-injection. To induce GBM-like tumors, we have introduced gene delivery and gene editing reagents to alter six major human GBM-associated signaling pathways in a cohort of swine. Resulting tumors will be examined molecularly to detect the pathway-associated transposons and CRISPR/Cas9-induced mutations in tumor tissue and determine the resemblance to human GBM. This somatic cell gene-modification platform will be adaptable, allowing on-demand inclusion of preclinical study-relevant alterations in GBM tumors, and will allow the rapid development tumors histologically and genetically similar to human GBM, which will be valuable for use in pre-clinical therapeutic studies, imaging studies using human clinical grade equipment, and surgical technique development.

scholarly journals TMOD-20. A SWINE MODEL OF GLIOBLASTOMA INDUCED BY SOMATIC GENE MODIFICATION

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii232-ii232
Author(s):  
Barbara Tschida ◽  
Dylan Duerre ◽  
Mandy Taisto ◽  
Adrienne Watson
Keyword(s):  
Brain Size ◽  
Expression Vectors ◽  
Efficient Production ◽  
Swine Model ◽  
Large Animal ◽  
Gene Modification ◽  
Large Animal Models ◽  
Therapeutic Development ◽  
Human Gbm

Abstract Glioblastoma (GBM) is the most common and malignant primary brain tumor. Novel therapeutic development for GBM is needed since the standard of care universally fails to cure patients and the five-year survival rate remains below 10%. GBM therapeutic development is hampered by the lack of relevant large animal models for preclinical studies. To mitigate this problem, we are developing a model of GBM in outbred, immune-proficient swine which have comparable brain size and anatomy to humans. We developed methods for introducing genome engineering tools to minipig brain cells in vivo by direct injection of gene delivery reagents to the lateral ventricle. Using this technique, we have delivered a combination of expression vectors for oncogenes and targeted nucleases to disrupt tumor suppressor genes commonly altered in human GBM to alter six major human GBM-associated signaling pathways in a cohort of minipigs (Ras, Pi3k, p53, Rb/E2F, Pdgf, and the alternative lengthening of telomeres (ALT) pathways). These minipigs are being monitored for tumorigenesis using a secreted reporter, detectable through a simple luminescence-based blood test. Resulting tumors will be examined molecularly to detect the pathway-associated alterations in tumor tissue and determine the resemblance to human GBM. We hypothesize that this somatic cell gene-modification platform we have developed in the minipig will facilitate the efficient production of brain tumors that histologically and genetically resemble human GBM. It will allow the production of tumors that are genetically heterogeneous, of specified molecular subclasses, containing therapeutic targets of interest, and in the context of genetic backgrounds of interest. This minipig model of GBM will be applied towards preclinical therapeutic studies, imaging studies using human clinical grade equipment, and surgical technique development, to improve clinical trial success rates and patient outcomes. Funding for this study is provided by the National Institutes of Health through SBIR grant # 1R43CA235837-01A1.

scholarly journals Parameters of biliary hydrodynamic injection during endoscopic retrograde cholangio-pancreatography in pigs for applications in gene delivery

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0249931
Author(s):  
Yuting Huang ◽  
Robert L. Kruse ◽  
Hui Ding ◽  
Mohamad I. Itani ◽  
Jonathan Morrison ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Vital Signs ◽  
Biliary Tree ◽  
Large Animal ◽  
Endoscopic Retrograde ◽  
Flow Rates ◽  
Hydrodynamic Injection ◽  
Large Animal Models ◽  
Injection Parameters ◽  
Tree Injury

The biliary system is routinely accessed for clinical purposes via endoscopic retrograde cholangiopancreatography (ERCP). We previously pioneered ERCP-mediated hydrodynamic injection in large animal models as an innovative gene delivery approach for monogenic liver diseases. However, the procedure poses potential safety concerns related mainly to liver or biliary tree injury. Here, we sought to further define biliary hydrodynamic injection parameters that are well-tolerated in a human-sized animal model. ERCP was performed in pigs, and hydrodynamic injection carried out using a novel protocol to reduce duct wall stress. Each pig was subjected to multiple repeated injections to expedite testing and judge tolerability. Different injection parameters (volume, flow rate) and injection port diameters were tested. Vital signs were monitored throughout the procedure, and liver enzyme panels were collected pre- and post-procedure. Pigs tolerated repeated biliary hydrodynamic injections with only occasional, mild, isolated elevation in aspartate aminotransferase (AST), which returned to normal levels within one day post-injection. All other liver tests remained unchanged. No upper limit of volume tolerance was reached, which suggests the biliary tree can readily transmit fluid into the vascular space. Flow rates up to 10 mL/sec were also tolerated with minimal disturbance to vital signs and no anatomic rupture of bile ducts. Measured intrabiliary pressure was up to 150 mmHg, and fluid-filled vesicles were induced in liver histology at high flow rates, mimicking the changes in histology observed in mouse liver after hydrodynamic tail vein injection. Overall, our investigations in a human-sized pig liver using standard clinical equipment suggest that ERCP-guided hydrodynamic injection will be safely tolerated in patients. Future investigations will interrogate if higher flow rates and pressure mediate higher DNA delivery efficiencies.

A Review of Stem Cell Therapy for Spinal Cord Injury: Large Animal Models and the Frontier in Humans

2017 ◽  
Vol 98 ◽  
pp. 438-443 ◽  
Author(s):  
Brandon C. Gabel ◽  
Erik I. Curtis ◽  
Martin Marsala ◽  
Joseph D. Ciacci
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Stem Cell ◽  
Animal Models ◽  
Cell Therapy ◽  
Stem Cell Therapy ◽  
Large Animal ◽  
Large Animal Models ◽  
Cord Injury

scholarly journals Motor unit number index detects the effectiveness of surgical treatment in improving distal motor neuron loss in patients with incomplete cervical spinal cord injury

2020 ◽  
Author(s):  
Jun Li ◽  
Yancheng Zhu ◽  
Yang Li ◽  
ShiSheng He ◽  
Deguo Wang
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Surgical Treatment ◽  
Muscle Strength ◽  
Motor Neurons ◽  
Surgical Intervention ◽  
Cervical Spinal Cord ◽  
Injury Site ◽  
Cord Injury ◽  
Motor Neuron Loss

Abstract Background Recovery of motor dysfunction is important for patients with incomplete cervical spinal cord injury (SCI). To enhance the recovery of muscle strength, both research and treatments mainly focus on injury of upper motor neurons at the direct injury site. However, accumulating evidences have suggested that SCI has a downstream effect on the peripheral nervous system, which may contribute to the poor improvement of the muscle strength after operation. The aim of this study is to investigate the impact of early vs. delayed surgical intervention on the lower motor neurons (LMNs) distal to the injury site in patients with incomplete cervical SCI. Methods Motor unit number index (MUNIX) was performed on the tibialis anterior (TA), extensor digitorum brevis (EDB) and abductor hallucis (AH) in 47 patients with incomplete cervical SCI (early vs. delayed surgical-treatment: 17 vs. 30) and 34 healthy subjects approximately 12 months after operation. All patients were further assessed by American spinal injury association (ASIA) motor scales and Medical Research Council (MRC) scales. Results There are no difference of both ASIA motor scores and MRC scales between the patients who accepted early and delayed surgical treatment (P > 0.05). In contrast, the patients undergoing early surgical treatment showed lower MUSIX values in both bilateral EDB and bilateral TA, along with greater MUNIX values in both right-side EDB and right-side TA, compared to the patients who accepted delayed surgical treatment (P < 0.05). Conclusions Cervical SCI has a negative effect on the LMNs distal to the injury site. Early surgical intervention in Cervical SCI patients may improve the dysfunction of LMNs distal to the injury site, reducing secondary motor neuron loss, and eventually improving clinical prognosis.

scholarly journals Human Motor Neuron Progenitor Transplantation Leads to Endogenous Neuronal Sparing in 3 Models of Motor Neuron Loss

2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
Tanya J. Wyatt ◽  
Sharyn L. Rossi ◽  
Monica M. Siegenthaler ◽  
Jennifer Frame ◽  
Rockelle Robles ◽  
...  
Keyword(s):  
Animal Models ◽  
Motor Neuron ◽  
Motor Neurons ◽  
Embryonic Stem ◽  
Active Growth ◽  
Neuron Loss ◽  
Cord Injury ◽  
Human Motor ◽  
Motor Neuron Loss

Motor neuron loss is characteristic of many neurodegenerative disorders and results in rapid loss of muscle control, paralysis, and eventual death in severe cases. In order to investigate the neurotrophic effects of a motor neuron lineage graft, we transplanted human embryonic stem cell-derived motor neuron progenitors (hMNPs) and examined their histopathological effect in three animal models of motor neuron loss. Specifically, we transplanted hMNPs into rodent models of SMA (Δ7SMN), ALS (SOD1 G93A), and spinal cord injury (SCI). The transplanted cells survived and differentiated in all models. In addition, we have also found that hMNPs secrete physiologically active growth factorsin vivo, including NGF and NT-3, which significantly enhanced the number of spared endogenous neurons in all three animal models. The ability to maintain dying motor neurons by delivering motor neuron-specific neurotrophic support represents a powerful treatment strategy for diseases characterized by motor neuron loss.

scholarly journals Atorvastatin Inhibits Myocardial Apoptosis in a Swine Model of Coronary Microembolization by Regulating PTEN/PI3K/Akt Signaling Pathway

2016 ◽  
Vol 38 (1) ◽  
pp. 207-219 ◽  
Author(s):  
Jiangyou Wang ◽  
Han Chen ◽  
You Zhou ◽  
Qiang Su ◽  
Tao Liu ◽  
...  
Keyword(s):  
Cardiac Function ◽  
Tunel Staining ◽  
Swine Model ◽  
Large Animal ◽  
Functional Protein ◽  
Coronary Microembolization ◽  
Beneficial Effects ◽  
Large Animal Models ◽  
Myocardial Apoptosis

Background/Aims: Phosphatase and tensin homolog deleted on chromosome ten (PTEN) has been recognized as a promoter of apoptosis in various tissues, and revealed to be up-regulated in circumstances of coronary microembolization (CME). However, whether this functional protein could be modified by pretreatment of atorvastatin in models of CME has not been disclosed yet. Methods: Swine CME was induced by intra-coronary injection of inertia plastic microspheres (diameter 42 μm) into left anterior descending coronary, with or without pretreatment of atorvastatin or PTEN siRNA. Echocardiologic measurements, pathologic examination, TUNEL staining and western blotting were applied to assess their functional, morphological and molecular effects in CME. Results: PTEN were aberrantly up-regulated in cardiomyocytes following CME, with both the mRNA and protein levels increased after CME modeling. Pretreatment with atorvastatin could attenuate the induction of PTEN. Furthermore, down-regulation of PTEN in vivo via siRNA was associated with an improved cardiac function, attenuated myocardial apoptosis, and concomitantly inhibited expressions of key proapoptotic proteins such as Bax, cleaved-caspase-3. Interestingly, atorvastatin could markedly attenuate PTEN expression and therefore partially reverse cardiac dysfunction and attenuate the apoptosis of the myocardium following CME. Conclusion: Modulation of PTEN was probably as a potential mechanism involved in the beneficial effects of pretreatment of atorvastatin to cardiac function and apoptosis in large animal models of CME.

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