Publications by authors named "Hiroshi Nakagawa"

415 Publications

Super and Non-responders To Catheter Ablation for Atrial Fibrillation: A Quality-of-Life Assessment Using Patient Reported Outcomes.

Circ Arrhythm Electrophysiol 2021 Jul 19. Epub 2021 Jul 19.

Heart and Vascular Institute, Cleveland Clinic, Cleveland, OH.

- Atrial fibrillation (AF) ablation targets improvement in quality of life (QoL). Data is scarce on predictors of QoL improvement following ablation. We aimed to investigate the clinical characteristics underlying differential response in QoL after AF ablation (with or without arrhythmia recurrence). - All patients undergoing AF ablation (2013-2016) at our center were prospectively enrolled in a fully automated patient-reported outcomes registry. A large number of variables were collected including AF symptom severity scale (AFSSS) and AF burden (as indicated by AF frequency and duration scores). Patients were divided into 3 groups based on self-report of QoL improvement: remarkable (super responders), mild/moderate, and unchanged or worse (non-responders). Univariable and multivariable logistic regression models assessed clinical characteristics and QoL outcomes. - A total of 956 patients were included (25% females, mean age 63.9). Most patients (~80%) were super responders (n=761), 138 (14.4%) had mild/moderate improvement, and 57 (5.9%) were non-responders. The strongest predictors of remarkable QoL improvement were freedom of arrhythmia recurrence (OR 2.42, 95% CI 1.27-4.59, p-value = 0.007), and lower AF burden at 12 months. Similarly, higher AF burden was significantly associated with clinical "non-response". In patients with observed clinical recurrence-QoL mismatch, changes in AF burden at 12 months were the main predictors of QoL outcome, with lower burden predicting higher improvement in QoL and vice versa. - Most patients derive significant QoL benefit from AF ablation. Freedom from arrhythmia recurrence and lower AF burden predict remarkable QoL improvement following ablation.
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http://dx.doi.org/10.1161/CIRCEP.121.009938DOI Listing
July 2021

Conformational dynamics of a multi-domain protein by neutron scattering and computational analysis.

Biophys J 2021 Jul 6. Epub 2021 Jul 6.

J-PARC Center,Japan Atomic Energy Agency, Tokai, Ibaraki 319-1195,Japan.

The flexible conformations of a multi-domain protein are responsible for its biological functions. Although MurD, a 47-kDa protein which consists of three domains, sequentially changes its domain conformation from an open form to a closed form through a semi-closed form in its enzymatic reaction, the domain dynamics in each conformation remains unclear. In this study, we verify the conformational dynamics of MurD in the corresponding three states (apo, ATP- and inhibitor-bound states), with a combination of small-angle X-ray and neutron scattering (SAXS and SANS), dynamic light scattering (DLS), neutron backscattering (NBS), neutron spin echo (NSE) spectroscopy and molecular dynamics (MD) simulations. Applying principal component analysis of the MD trajectories, a twisting and an open-closed domain modes are identified as the major collective coordinates. The deviations of the experimental SAXS profiles from the theoretical calculations based on the known crystal structures becomes smaller in the ATP-bound state than in the apo state, and a further decrease is evident upon inhibitor-binding. These results suggest that domain motions of the protein are suppressed step-by-step of each ligand binding. The DLS and NBS data yield collective- and self-translational diffusion constants, respectively, and we used them to extract collective domain motions in nanometer and nanosecond scale from the NSE data. In the apo state, MurD shows both twisting and open-closed domain modes, while an ATP binding suppresses twisting domain motions and a further reduction of open-closed mode is seen in the inhibitor binding state. These observations are consistent with the structure modifications measured by the small-angle scattering as well as the MD simulations. Such changes in the domain dynamics associated with the sequential enzymatic reactions should be related to the affinity and reaction efficiency with a ligand that binds specifically to each reaction state.
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http://dx.doi.org/10.1016/j.bpj.2021.07.001DOI Listing
July 2021

Conformational ensemble of a multidomain protein explored by Gd electron paramagnetic resonance.

Biophys J 2021 Jul 7. Epub 2021 Jul 7.

Graduate School of Chemical Sciences and Engineering, Hokkaido University, Sapporo, Japan; Department of Chemistry, Faculty of Science, Hokkaido University, Sapporo, Japan. Electronic address:

Despite their importance in function, the conformational state of proteins and its changes are often poorly understood, mainly because of the lack of an efficient tool. MurD, a 47-kDa protein enzyme responsible for peptidoglycan biosynthesis, is one of those proteins whose conformational states and changes during their catalytic cycle are not well understood. Although it has been considered that MurD takes a single conformational state in solution as shown by a crystal structure, the solution nuclear magnetic resonance (NMR) study suggested the existence of multiple conformational state of apo MurD in solution. However, the conformational distribution has not been evaluated. In this work, we investigate the conformational states of MurD by the use of electron paramagnetic resonance (EPR), especially intergadolinium distance measurement using double electron-electron resonance (DEER) measurement. The gadolinium ions are fixed on specific positions on MurD via a rigid double-arm paramagnetic lanthanide tag that has been originally developed for paramagnetic NMR. The combined use of NMR and EPR enables accurate interpretation of the DEER distance information to the structural information of MurD. The DEER distance measurement for apo MurD shows a broad distance distribution, whereas the presence of the inhibitor narrows the distance distribution. The results suggest that MurD exists in a wide variety of conformational states in the absence of ligands, whereas binding of the inhibitor eliminates variation in conformational states. The multiple conformational states of MurD were previously implied by NMR experiments, but our DEER data provided structural characterization of the conformational variety of MurD.
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http://dx.doi.org/10.1016/j.bpj.2021.06.033DOI Listing
July 2021

Stem cells and origins of cancer in the upper gastrointestinal tract.

Cell Stem Cell 2021 Jun 10. Epub 2021 Jun 10.

Division of Digestive and Liver Diseases, Department of Medicine, Columbia University, College of Physicians and Surgeons, 1130 St. Nicholas Avenue, New York, NY 10032, USA; Herbert Irving Comprehensive Cancer Center, 1130 St. Nicholas Avenue, New York, NY 10032, USA. Electronic address:

The esophagus and stomach, joined by a unique transitional zone, contain actively dividing epithelial stem cells required for organ homeostasis. Upon prolonged inflammation, epithelial cells in both organs can undergo a cell fate switch leading to intestinal metaplasia, predisposing to malignancy. Here we discuss the biology of gastroesophageal stem cells and their role as cells of origin in cancer. We summarize the interactions between the stromal niche and gastroesophageal stem cells in metaplasia and early expansion of mutated stem-cell-derived clones during carcinogenesis. Finally, we review new approaches under development to better study gastroesophageal stem cells and advance the field.
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http://dx.doi.org/10.1016/j.stem.2021.05.012DOI Listing
June 2021

Effect of trivalent chromium on erythropoietin production and the prevention of insulin resistance in HepG2 cells.

Arch Biochem Biophys 2021 Sep 5;708:108960. Epub 2021 Jun 5.

Laboratory of Bioenvironmental Sciences, Course of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-58 Rinku Ohrai-Kita, Izumisano, Osaka, 598-8531, Japan.

In erythropoietin (EPO)-producing HepG2 cells, we investigated the effect of trivalent chromium (Cr) on the promotion of EPO production and the induction of insulin resistance. Cr increased hypoxia-inducible factor (HIF)-1α protein, EPO mRNA expression and EPO protein levels in HepG2 cells. The effect of Cr on EPO production was inhibited by inhibition of proliferator-activated receptor γ (PPARγ). Insulin resistance was induced by culturing with insulin resistance induction medium supplemented with palmitic acid for 24 h. When Cr was added to the medium, the increase in glucose-6-phosphatase and phosphoenolpyruvate carboxykinase 1 mRNA expression levels and the decrease in the ratio of phosphorylated Akt to Akt protein were suppressed, and the induction of insulin resistance prevented. When a PPARγ inhibitor or siPPARγ was added together with Cr, the inhibitory effect of Cr on the induction of insulin resistance disappeared. In addition, pretreatment with siEPO suppressed the increase in EPO mRNA expression, and the inhibitory effect on the induction of insulin resistance due to the addition of Cr was significantly reduced. These results suggest that the inhibition of insulin resistance induction by Cr in HepG2 cells involves the promotion of EPO production mediated by PPARγ, in addition to other PPARγ-mediated activities.
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http://dx.doi.org/10.1016/j.abb.2021.108960DOI Listing
September 2021

An enhanced therapeutic effect of repetitive transcranial magnetic stimulation combined with antibody treatment in a primate model of spinal cord injury.

PLoS One 2021 2;16(6):e0252023. Epub 2021 Jun 2.

Systems Neuroscience Section, Department of Neuroscience, Primate Research Institute, Kyoto University, Inuyama, Aichi, Japan.

Repetitive transcranial magnetic stimulation (rTMS) targeting the primary motor cortex (MI) is expected to provide a therapeutic impact on spinal cord injury (SCI). On the other hand, treatment with antibody against repulsive guidance molecule-a (RGMa) has been shown to ameliorate motor deficits after SCI in rodents and primates. Facilitating activity of the corticospinal tract (CST) by rTMS following rewiring of CST fibers by anti-RGMa antibody treatment may exert an enhanced effect on motor recovery in a primate model of SCI. To address this issue, we examined whether such a combined therapeutic strategy could contribute to accelerating functional restoration from SCI. In our SCI model, unilateral lesions were made between the C6 and the C7 level. Two macaque monkeys were used for each of the combined therapy and antibody treatment alone, while one monkey was for rTMS alone. The antibody treatment was continuously carried out for four weeks immediately after SCI, and rTMS trials applying a thermoplastic mask and a laser distance meter lasted ten weeks. Behavioral assessment was performed over 14 weeks after SCI to investigate the extent to which motor functions were restored with the antibody treatment and/or rTMS. While rTMS without the preceding antibody treatment produced no discernible sign for functional recovery, a combination of the antibody and rTMS exhibited a greater effect, especially at an early stage of rTMS trials, on restoration of dexterous hand movements. The present results indicate that rTMS combined with anti-RGMa antibody treatment may exert a synergistic effect on motor recovery from SCI.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0252023PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8172028PMC
June 2021

NOTCH3 limits the epithelial-mesenchymal transition and predicts a favorable clinical outcome in esophageal cancer.

Cancer Med 2021 Jun 27;10(12):3986-3996. Epub 2021 May 27.

Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.

Background: Esophageal squamous cell carcinoma (ESCC) is the deadliest of all human squamous cell carcinomas and is characterized by chemotherapy resistance and poor prognosis associated with the epithelial-mesenchymal transition (EMT). A subset of ESCC displays loss-of-function mutations in genes encoding Notch receptor family members, including NOTCH3. Although Notch signaling regulates EMT in ESCC cells, the role of NOTCH3 in EMT and chemotherapy resistance remains elusive. This study aimed to examine the role of NOTCH3 in EMT and chemotherapy resistance, and determine whether NOTCH3 expression can be used to predict the response to chemotherapy.

Methods: In vitro and in vivo assays were conducted to clarify the contribution of NOTCH3 to chemotherapy resistance. Using specimens from 120 ESCC patients treated with neoadjuvant chemotherapy, we compared the expression levels of NOTCH3 and genes involved in EMT according to the degree of chemotherapy sensitivity.

Results: In ESCC cells, chemotherapy resistance was associated with NOTCH3 downregulation and concurrent activation of EMT. RNA interference to silence NOTCH3 resulted in induction of the EMT marker Vimentin (VIM), leading to chemotherapy resistance in ESCC cells. Conversely, ectopic expression of the activated form of NOTCH3 suppressed EMT and sensitized cells to chemotherapy. Results of chromatin immunoprecipitation assays suggested that NOTCH3 may repress transcription of the VIM.

Conclusions: Our findings suggest that NOTCH3 may control chemotherapy sensitivity by regulating EMT. NOTCH3 may serve as a novel biomarker to predict better clinical outcomes in ESCC patients.
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http://dx.doi.org/10.1002/cam4.3933DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8209574PMC
June 2021

Reprogramming of the esophageal squamous carcinoma epigenome by SOX2 promotes ADAR1 dependence.

Nat Genet 2021 06 10;53(6):881-894. Epub 2021 May 10.

Herbert Irving Comprehensive Cancer Center, Division of Digestive and Liver Diseases, Department of Medicine, Columbia University, New York, NY, USA.

Esophageal squamous cell carcinomas (ESCCs) harbor recurrent chromosome 3q amplifications that target the transcription factor SOX2. Beyond its role as an oncogene in ESCC, SOX2 acts in development of the squamous esophagus and maintenance of adult esophageal precursor cells. To compare Sox2 activity in normal and malignant tissue, we developed engineered murine esophageal organoids spanning normal esophagus to Sox2-induced squamous cell carcinoma and mapped Sox2 binding and the epigenetic and transcriptional landscape with evolution from normal to cancer. While oncogenic Sox2 largely maintains actions observed in normal tissue, Sox2 overexpression with p53 and p16 inactivation promotes chromatin remodeling and evolution of the Sox2 cistrome. With Klf5, oncogenic Sox2 acquires new binding sites and enhances activity of oncogenes such as Stat3. Moreover, oncogenic Sox2 activates endogenous retroviruses, inducing expression of double-stranded RNA and dependence on the RNA editing enzyme ADAR1. These data reveal SOX2 functions in ESCC, defining targetable vulnerabilities.
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http://dx.doi.org/10.1038/s41588-021-00859-2DOI Listing
June 2021

The utility of a novel mapping algorithm utilizing vectors and global pattern of propagation for scar-related atrial tachycardias.

J Cardiovasc Electrophysiol 2021 Jul 22;32(7):1909-1917. Epub 2021 May 22.

Section of Cardiac Pacing and Electrophysiology, Heart and Vascular Institute, Cleveland Clinic, Cleveland, Ohio, USA.

Background: Activation maps of scar-related atrial tachycardias (AT) can be challenging to interpret due to difficulty in inaccurate annotation of electrograms, and an arbitrarily predefined mapping window. A novel mapping software integrating vector data and applying an algorithmic solution taking into consideration global activation pattern has been recently described (Coherent™, Biosense Webster "Investigational").

Objective: We aimed to assess the investigational algorithm to determine the mechanism of AT compared with the standard algorithm.

Methods: This study included patients who underwent ablation of scar-related AT using the Carto 3 and the standard activation algorithm. The mapping data were analyzed retrospectively using the investigational algorithm, and the mechanisms were evaluated by two independent electrophysiologists.

Results: A total of 77 scar-related AT activation maps were analyzed (89.6% left atrium, median tachycardia cycle length of 273 ms). Of those, 67 cases with a confirmed mechanism of arrhythmia were used to compare the activation software. The actual mechanism of the arrhythmia was more likely to be identified with the investigational algorithm (67.2% vs. 44.8%, p = .009). In five patients with dual-loop circuits, 3/5 (60%) were correctly identified by the investigational algorithm compared to 0/5 (0%) with the standard software. The reduced atrial voltage was prone to lead to less capable identification of mechanism (p for trend: .05). The investigational algorithm showed higher inter-reviewer agreement (Cohen's kappa .62 vs. .47).

Conclusions: In patients with scar-related ATs, activation mapping algorithms integrating vector data and "best-fit" propagation solution may help in identifying the mechanism and the successful site of termination.
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http://dx.doi.org/10.1111/jce.15074DOI Listing
July 2021

Understanding the cellular origin and progression of esophageal cancer using esophageal organoids.

Cancer Lett 2021 Jul 7;509:39-52. Epub 2021 Apr 7.

Division of Digestive and Liver Diseases, Department of Medicine, Columbia University, New York, NY, USA; Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY, USA. Electronic address:

Three-dimensional (3D) organoids are a novel tool to model epithelial cell biology and human diseases of the esophagus. 3D organoid culture systems have been utilized to investigate the pathobiology of esophageal cancer, including both squamous cell carcinoma and adenocarcinoma. Additional organoid-based approaches for study of esophageal development and benign esophageal diseases have provided key insights into esophageal keratinocyte differentiation and mucosal regeneration. These investigations have implications for the identification of esophageal cancer stem cells, as well as the potential to halt malignant progression through induction of differentiation pathways. Patient-derived organoids (PDOs) from human tissue samples allow for unique and faithful in vitro modeling of esophageal cancers, and provide an exciting platform for investigation into personalized medicine and targeted treatment approaches, as well as new models for understanding therapy resistance and recurrent disease. Future directions include high-throughput genomic screening using PDOs, and study of tumor-microenvironmental interactions through co-culture with immune and stromal cells and novel extracellular matrix complexes.
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http://dx.doi.org/10.1016/j.canlet.2021.03.031DOI Listing
July 2021

Pan-ERBB kinase inhibition augments CDK4/6 inhibitor efficacy in oesophageal squamous cell carcinoma.

Gut 2021 Mar 31. Epub 2021 Mar 31.

Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA

Objective: Oesophageal squamous cell carcinoma (OSCC), like other squamous carcinomas, harbour highly recurrent cell cycle pathway alterations, especially hyperactivation of the CCND1/CDK4/6 axis, raising the potential for use of existing CDK4/6 inhibitors in these cancers. Although CDK4/6 inhibition has shown striking success when combined with endocrine therapy in oestrogen receptor positive breast cancer, CDK4/6 inhibitor palbociclib monotherapy has not revealed evidence of efficacy to date in OSCC clinical studies. Herein, we sought to elucidate the identification of key dependencies in OSCC as a foundation for the selection of targets whose blockade could be combined with CDK4/6 inhibition.

Design: We combined large-scale genomic dependency and pharmaceutical screening datasets with preclinical cell line models, to identified potential combination therapies in squamous cell cancer.

Results: We identified sensitivity to inhibitors to the ERBB family of receptor kinases, results clearly extending beyond the previously described minority of tumours with EGFR amplification/dependence, specifically finding a subset of OSCCs with dual dependence on ERBB3 and ERBB2. Subsequently. we demonstrated marked efficacy of combined pan-ERBB and CDK4/6 inhibition in vitro and in vivo. Furthermore, we demonstrated that squamous lineage transcription factor KLF5 facilitated activation of ERBBs in OSCC.

Conclusion: These results provide clear rationale for development of combined ERBB and CDK4/6 inhibition in these cancers and raises the potential for KLF5 expression as a candidate biomarker to guide the use of these agents. These data suggested that by combining existing Food and Drug Administration (FDA)-approved agents, we have the capacity to improve therapy for OSCC and other squamous cancer.
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http://dx.doi.org/10.1136/gutjnl-2020-323276DOI Listing
March 2021

Mutant p53 regulates Survivin to foster lung metastasis.

Genes Dev 2021 Apr 18;35(7-8):528-541. Epub 2021 Mar 18.

Herbert Irving Comprehensive Cancer Center, Columbia University, New York, New York 10032, USA.

Esophageal squamous cell carcinoma (ESCC) is one of the most lethal cancers worldwide and evolves often to lung metastasis. (homologous to in mice) is a common hot spot mutation. How metastasis is regulated by p53 in ESCC remains to be investigated. To investigate p53-mediated molecular mechanisms, we used a carcinogen-induced approach in mice to model ESCC. In the primary tumor cell lines, we depleted Trp53 (shTrp53) and observed a marked reduction in cell invasion in vitro and lung metastasis burden in a tail-vein injection model in comparing isogenic cells (shCtrl). Furthermore, we performed bulk RNA-seq to compare gene expression profiles of metastatic and primary shCtrl and shTrp53 cells. We identified the YAP- axis as a potential mediator of -mediated metastasis. We demonstrate that expression of Survivin, an antiapoptotic protein encoded by , increases in the presence of Trp53 Furthermore, depletion of Survivin specifically decreases Trp53-driven lung metastasis. Mechanistically, Trp53 but not wild-type Trp53, binds with YAP in ESCC cells, suggesting their cooperation to induce Survivin expression. Furthermore, Survivin high expression level is associated with increased metastasis in several GI cancers. Taken together, this study unravels new insights into how mutant p53 mediates metastasis.
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http://dx.doi.org/10.1101/gad.340505.120DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8015716PMC
April 2021

Association Between Earwax-Determinant Genotypes and Acquired Middle Ear Cholesteatoma in a Japanese Population.

Otolaryngol Head Neck Surg 2021 Mar 16:1945998211000374. Epub 2021 Mar 16.

Department of Otorhinolaryngology, Faculty of Medicine, Juntendo University, Tokyo, Japan.

Objective: A single-nucleotide polymorphism 538G>A in the human gene is a determinant of the earwax morphotype. 538GG and GA correspond to wet earwax and 538AA to dry earwax. Despite a putative positive correlation between the frequency of the 538G allele and the prevalence of cholesteatoma, minimal clinical information is currently available. We aimed to evaluate this association between the genotypes and acquired middle ear cholesteatoma.

Study Design: Case-control study.

Setting: Single-center academic hospital.

Methods: We recruited 67 Japanese patients with acquired middle ear cholesteatoma (cholesteatoma group) and 100 Japanese controls with no history of middle ear cholesteatoma. We assessed the genotypes for all participants. Clinical information was collected from the cholesteatoma group. The genotype data of 104 Japanese people from the 1000 Genomes Project who represent the general population were used.

Results: The proportion of participants with 538GG or GA was significantly higher in the cholesteatoma group than in the control group or general Japanese population ( < .001). The 538G allele frequency was also significantly higher in the cholesteatoma group than in the control group or general Japanese population ( < .001). Multivariate logistic regression analyses revealed a significant association between the genotype and acquired middle ear cholesteatoma (odds ratio, 5.49; 95% CI, 2.61-11.5; < .001).

Conclusion: Our results suggest that the genotypes could be associated with the development of acquired middle ear cholesteatoma among Japanese people.
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http://dx.doi.org/10.1177/01945998211000374DOI Listing
March 2021

Freezable and Unfreezable Hydration Water: Distinct Contributions to Protein Dynamics Revealed by Neutron Scattering.

J Phys Chem Lett 2021 Mar 25;12(8):2172-2176. Epub 2021 Feb 25.

Division of Biophysics, Department of Physiology, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.

Hydration water plays a crucial role for activating the protein dynamics required for functional expression. Yet, the details are not understood about how hydration water couples with protein dynamics. A temperature hysteresis of the ice formation of hydration water is a key phenomenon to understand which type of hydration water, unfreezable or freezable hydration water, is crucial for the activation of protein dynamics. Using neutron scattering, we observed a temperature-hysteresis phenomenon in the diffraction peaks of the ice of freezable hydration water, whereas protein dynamics did not show any temperature hysteresis. These results show that the protein dynamics is not coupled with freezable hydration water dynamics, and unfreezable hydration water is essential for the activation of protein dynamics. Decoupling of the dynamics between unfreezable and freezable hydration water could be the cause of the distinct contributions of hydration water to protein dynamics.
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http://dx.doi.org/10.1021/acs.jpclett.0c03786DOI Listing
March 2021

Morphological features of large layer V pyramidal neurons in cortical motor-related areas of macaque monkeys: analysis of basal dendrites.

Sci Rep 2021 Feb 18;11(1):4171. Epub 2021 Feb 18.

Systems Neuroscience Section, Primate Research Institute, Kyoto University, Inuyama, Aichi, 484-8506, Japan.

In primates, large layer V pyramidal neurons located in the frontal motor-related areas send a variety of motor commands to the spinal cord, giving rise to the corticospinal tract, for execution of skilled motor behavior. However, little is known about the morphological diversity of such pyramidal neurons among the areas. Here we show that the structure of basal dendrites of the large layer V pyramidal neurons in the dorsal premotor cortex (PMd) is different from those in the other areas, including the primary motor cortex, the supplementary motor area, and the ventral premotor cortex. In the PMd, not only the complexity (arborization) of basal dendrites, i.e., total dendritic length and branching number, was poorly developed, but also the density of dendritic spines was so low, as compared to the other motor-related areas. Regarding the distribution of the three dendritic spine types identified, we found that thin-type (more immature) spines were prominent in the PMd in comparison with stubby- and mushroom-type (more mature) spines, while both thin- and stubby-type spines were in the other areas. The differential morphological features of basal dendrites might reflect distinct patterns of motor information processing within the large layer V pyramidal neurons in individual motor-related areas.
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http://dx.doi.org/10.1038/s41598-021-83680-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7893167PMC
February 2021

Systematic Evaluation of High-Resolution Activation Mapping to Identify Residual Endocardial and Epicardial Conduction Across the Mitral Isthmus.

JACC Clin Electrophysiol 2021 03 27;7(3):292-304. Epub 2021 Jan 27.

Department of Cardiovascular Medicine, Cleveland Clinic, Cleveland, Ohio, USA. Electronic address:

Objectives: This study sought to systematically evaluate the ability of a high-resolution mapping system (Rhythmia, Boston Scientific, Marlborough, Massachusetts) to rapidly and accurately localize residual endocardial and epicardial conduction after mitral isthmus (MI) ablation, facilitating MI block.

Background: Achieving conduction block across the mitral isthmus (MI) is challenging.

Methods: Fifty consecutive patients undergoing MI ablation after pulmonary vein isolation were enrolled. After initial endocardial radiofrequency (RF) ablation across the lateral MI, high-resolution activation mapping of the MI with simultaneous coronary sinus (CS) mapping was performed to verify block or localize residual conduction across the MI during left atrial (LA) appendage and CS pacing. Propagation maps were used to identify residual conduction across the MI as endocardial, via the CS or Marshall tract.

Results: In all 50 patients, after the initial endocardial ablation across the MI, repeat high-resolution mapping of the LA and CS was obtained (median: 3,329 mapped points; 4.0 min of mapping time). The initial endocardial MI ablation resulted in block in 9 of 50 patients (18%). In the remaining 41 patients, the propagation map identified residual conduction in 4 patterns: 1) only endocardial gap in 12 patients (29%); 2) only CS connection in 10 patients (24%); 3) both endocardial and CS connections in 14 patients (34%); and 4) Marshall tract connection in 5 patients (12%). In 8 patients, the propagation map revealed residual conduction, despite differential atrial pacing suggesting bidirectional block. Focal ablation at the identified residual conduction site (median: 0.7 min of RF) resulted in block in 49 of 50 (98%) patients.

Conclusions: High-resolution propagation maps of the LA/CS rapidly and accurately localize residual endocardial and epicardial conduction across the MI. Focal ablation with short RF time at the identified gap(s) achieved complete block across MI in 98% of cases.
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http://dx.doi.org/10.1016/j.jacep.2020.09.025DOI Listing
March 2021

MicroRNA-Based Cancer Mortality Risk Scoring System and hTERT Expression in Early-Stage Oral Squamous Cell Carcinoma.

J Oncol 2021 15;2021:8292453. Epub 2021 Jan 15.

Columbia University Irving Medical Center, New York, NY, USA.

We have previously constructed a novel microRNA (miRNA)-based prognostic model and cancer-specific mortality risk score formula to predict survival outcome in oral squamous cell carcinoma (OSCC) patients who are already categorized into "early-stage" by the TNM staging system. A total of 836 early-stage OSCC patients were assigned the mortality risk scores. We evaluated the efficacy of various treatment regimens in terms of survival benefit compared to surgery only in patients stratified into high (risk score ≥0) versus low (risk score <0) mortality risk categories. For the high-risk group, surgery with neck dissection significantly improved the 5-year survival to 75% from 46% with surgery only ( < 0.001); a Cox proportional hazard model on time-to-death demonstrated a hazard ratio of 0.37 for surgery with neck dissection (95% CI: 0.2-0.6; =0.0005). For the low-risk group, surgery only was the treatment of choice associated with 5-year survival benefit. Regardless of treatment selected, those with risk score ≥2 may benefit from additional therapy to prevent cancer relapse. We also identified hTERT (human telomerase reverse transcriptase) as a gene target common to the prognostic miRNAs. There was 22-fold increase in the hTERT expression level in patients with risk score ≥2 compared to healthy controls ( < 0.0005). Overexpression of hTERT was also observed in the patient-derived OSCC organoid compared to that of normal organoid. The DNA cancer vaccine that targets hTERT-expressing cells currently undergoing rigorous clinical evaluation for other tumors can be repurposed to prevent cancer recurrence in these high-risk early-stage oral cancer patients.
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http://dx.doi.org/10.1155/2021/8292453DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7822680PMC
January 2021

Determining the effect of water temperature on the T1 and T2 relaxation times of the lung tissue at 9.4 T MRI: A drowning mouse model.

Leg Med (Tokyo) 2021 Mar 4;49:101836. Epub 2021 Jan 4.

Department of Forensic Medicine, The Jikei University School of Medicine, Tokyo, Japan. Electronic address:

Japanese individuals have a unique culture of soaking in a bathtub, and forensic pathologists have experienced fatal cases due to drowning. However, T1 and T2 relaxation times of a drowning lung are poorly documented. In the present study, we investigated the relationship between drowning water temperature and T1 and T2 relaxation times of drowning lung tissues at 9.4 T MRI (Bruker, BioSpec94/20USR). The mice used as animal drowning models were directly submerged in freshwater. Water temperature was set to 8 °C-10 °C (cold), 20 °C-22 °C (normal), 30 °C, and 45 °C. The regions of interest (ROIs) on the axial section of the third slice were set at the central and peripheral areas of each-the left and the right-lung. T1 relaxation times measured immediately after death differed by the presence or absence of soaking water, except in case of cold water temperature. In the drowning groups, T1 relaxation time showed a linear dependency on water temperature. By contrast, T2 relaxation time was almost constant regardless of the presence of drowning under the same temperature condition; when compared in the lung areas of the same individuals, the times were uniformly reduced in drowning models. To minimize the effects of hypostasis and decomposition, we performed measurements immediately after death and were able to determine the noticeable difference in drowning water temperature. These results may be useful for qualitative assessments of a drowning lung and may serve as a basis when imaging the human body during forensic autopsy cases.
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http://dx.doi.org/10.1016/j.legalmed.2020.101836DOI Listing
March 2021

Rab11-FIP1 mediates epithelial-mesenchymal transition and invasion in esophageal cancer.

EMBO Rep 2021 02 6;22(2):e48351. Epub 2021 Jan 6.

Herbert Irving Comprehensive Cancer Center, Division of Digestive and Liver Diseases, Department of Medicine, Columbia University, New York, NY, USA.

Esophageal squamous cell carcinoma (ESCC) is the most common subtype of esophageal cancer worldwide. The most commonly mutated gene in ESCC is TP53. Using a combinatorial genetic and carcinogenic approach, we generate a novel mouse model of ESCC expressing either mutant or null p53 and show that mutant p53 exhibits enhanced tumorigenic properties and displays a distinct genomic profile. Through RNA-seq analysis, we identify several endocytic recycling genes, including Rab Coupling Protein (Rab11-FIP1), which are significantly downregulated in mutant p53 tumor cells. In 3-dimensional (3D) organoid models, genetic knockdown of Rab11-FIP1 results in increased organoid size. Loss of Rab11-FIP1 increases tumor cell invasion in part through mutant p53 but also in an independent manner. Furthermore, loss of Rab11-FIP1 in human ESCC cell lines decreases E-cadherin expression and increases mesenchymal lineage-specific markers, suggesting induction of epithelial-mesenchymal transition (EMT). Rab11-FIP1 regulates EMT through direct inhibition of Zeb1, a key EMT transcriptional factor. Our novel findings reveal that Rab11-FIP1 regulates organoid formation, tumor cell invasion, and EMT.
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http://dx.doi.org/10.15252/embr.201948351DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7857540PMC
February 2021

Radiofrequency ablation using a needle electrode combined with heated saline injection: Three different mechanisms of tissue heating.

Heart Rhythm 2021 Mar 11;18(3):453-454. Epub 2020 Dec 11.

Department of Cardiovascular Medicine, Cleveland Clinic, Cleveland, Ohio.

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http://dx.doi.org/10.1016/j.hrthm.2020.12.008DOI Listing
March 2021

Dynamics of proteins with different molecular structures under solution condition.

Sci Rep 2020 12 10;10(1):21678. Epub 2020 Dec 10.

Institute for Integrated Radiation and Nuclear Science, Kyoto University, Kumatori, Sennan-gun, Osaka, 590-0494, Japan.

Incoherent quasielastic neutron scattering (iQENS) is a fascinating technique for investigating the internal dynamics of protein. However, low flux of neutron beam, low signal to noise ratio of QENS spectrometers and unavailability of well-established analyzing method have been obstacles for studying internal dynamics under physiological condition (in solution). The recent progress of neutron source and spectrometer provide the fine iQENS profile with high statistics and as well the progress of computational technique enable us to quantitatively reveal the internal dynamic from the obtained iQENS profile. The internal dynamics of two proteins, globular domain protein (GDP) and intrinsically disordered protein (IDP) in solution, were measured with the state-of-the art QENS spectrometer and then revealed with the newly developed analyzing method. It was clarified that the average relaxation rate of IDP was larger than that of GDP and the fraction of mobile H atoms of IDP was also much higher than that of GDP. Combined with the structural analysis and the calculation of solvent accessible surface area of amino acid residue, it was concluded that the internal dynamics were related to the highly solvent exposed amino acid residues depending upon protein's structure.
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http://dx.doi.org/10.1038/s41598-020-78311-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7728768PMC
December 2020

Carbon tetrachloride suppresses ER-Golgi transport by inhibiting COPII vesicle formation on the ER membrane in the RLC-16 hepatocyte cell line.

Cell Biol Int 2021 Mar 8;45(3):633-641. Epub 2020 Dec 8.

Laboratory of Toxicology, Graduate School of Veterinary Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.

Carbon tetrachloride (CCl ) causes hepatotoxicity in mammals, with its hepatocytic metabolism producing radicals that attack the intracellular membrane system and destabilize intracellular vesicle transport. Inhibition of intracellular transport causes lipid droplet retention and abnormal protein distribution. The intracellular transport of synthesized lipids and proteins from the endoplasmic reticulum (ER) to the Golgi apparatus is performed by coat complex II (COPII) vesicle transport, but how CCl inhibits COPII vesicle transport has not been elucidated. COPII vesicle formation on the ER membrane is initiated by the recruitment of Sar1 protein from the cytoplasm to the ER membrane, followed by that of the COPII coat constituent proteins (Sec23, Sec24, Sec13, and Sec31). In this study, we evaluated the effect of CCl on COPII vesicle formation using the RLC-16 rat hepatocyte cell line. Our results showed that CCl suppressed ER-Golgi transport in RLC-16 cells. Using a reconstituted system of rat liver tissue-derived cytoplasm and RLC-16 cell-derived ER membranes, CCl treatment inhibited the recruitment of Sar1 and Sec13 from the cytosolic fraction to ER membranes. CCl -induced changes in the ER membrane accordingly inhibited the accumulation of COPII vesicle-coated constituent proteins on the ER membrane, as well as the formation of COPII vesicles, which suppressed lipid and protein transport between the ER and Golgi apparatus. Our data suggest that CCl inhibits ER-Golgi intracellular transport by inhibiting COPII vesicle formation on the ER membrane in hepatocytes.
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http://dx.doi.org/10.1002/cbin.11510DOI Listing
March 2021

Mitochondrial dysfunction in inflammatory bowel disease alters intestinal epithelial metabolism of hepatic acylcarnitines.

J Clin Invest 2021 Jan;131(1)

Department of Medicine, Division of Gastroenterology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

As the interface between the gut microbiota and the mucosal immune system, there has been great interest in the maintenance of colonic epithelial integrity through mitochondrial oxidation of butyrate, a short-chain fatty acid produced by the gut microbiota. Herein, we showed that the intestinal epithelium could also oxidize long-chain fatty acids, and that luminally delivered acylcarnitines in bile could be consumed via apical absorption by the intestinal epithelium, resulting in mitochondrial oxidation. Finally, intestinal inflammation led to mitochondrial dysfunction in the apical domain of the surface epithelium that may reduce the consumption of fatty acids, contributing to higher concentrations of fecal acylcarnitines in murine Citrobacter rodentium-induced colitis and human inflammatory bowel disease. These results emphasized the importance of both the gut microbiota and the liver in the delivery of energy substrates for mitochondrial metabolism by the intestinal epithelium.
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http://dx.doi.org/10.1172/JCI133371DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7773399PMC
January 2021

Choosing the optimal population for a genome-wide association study: A simulation of whole-genome sequences from rice.

Plant Genome 2020 03 19;13(1):e20005. Epub 2020 Mar 19.

Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The Univ. of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.

A genome-wide association study (GWAS) needs to have a suitable population. The factors that affect a GWAS (e.g. population structure, sample size, and sequence analysis and field testing costs) need to be considered. Mixed populations containing subpopulations of different genetic backgrounds may be suitable populations. We conducted simulation experiments to see if a population with high genetic diversity, such as a diversity panel, should be added to a target population, especially when the target population harbors small genetic diversity. The target population was 112 accessions of Oryza sativa L. subsp. japonica, mainly developed in Japan. We combined the target population with three populations that had higher genetic diversity. These were 100 indica accessions, 100 japonica accessions, and 100 accessions with various genetic backgrounds. The results showed that the GWAS's power with a mixed population was generally higher than with a separate population. Also, the optimal GWAS populations varied depending on the fixation index (F ) of the quantitative trait nucleotides (QTNs) and the polymorphism of QTNs in each population. When a QTN was polymorphic in a target population, a target population combined with a higher diversity population improved the QTN's detection power. By investigating F and the expected heterozygosity (H ) as factors influencing the detection power, we showed that single nucleotide polymorphisms with high F or low H are less likely to be detected by GWAS with mixed populations. Sequenced or genotyped germplasm collections can improve the GWAS's detection power by using a subset of the collections with a target population.
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http://dx.doi.org/10.1002/tpg2.20005DOI Listing
March 2020

Autophagy mitigates ethanol-induced mitochondrial dysfunction and oxidative stress in esophageal keratinocytes.

PLoS One 2020 23;15(9):e0239625. Epub 2020 Sep 23.

Herbert Irving Comprehensive Cancer Center, Columbia University, New York, New York, United States of America.

During alcohol consumption, the esophageal mucosa is directly exposed to high concentrations of ethanol (EtOH). We therefore investigated the response of normal human esophageal epithelial cell lines EPC1, EPC2 and EPC3 to acute EtOH exposure. While these cells were able to tolerate 2% EtOH for 8 h in both three-dimensional organoids and monolayer culture conditions, RNA sequencing suggested that EtOH induced mitochondrial dysfunction. With EtOH treatment, EPC1 and EPC2 cells also demonstrated decreased mitochondrial ATPB protein expression by immunofluorescence and swollen mitochondria lacking intact cristae by transmission electron microscopy. Mitochondrial membrane potential (ΔΨm) was decreased in a subset of EPC1 and EPC2 cells stained with ΔΨm-sensitive dye MitoTracker Deep Red. In EPC2, EtOH decreased ATP level while impairing mitochondrial respiration and electron transportation chain functions, as determined by ATP fluorometric assay, respirometry, and liquid chromatography-mass spectrometry. Additionally, EPC2 cells demonstrated enhanced oxidative stress by flow cytometry for mitochondrial superoxide (MitoSOX), which was antagonized by the mitochondria-specific antioxidant MitoCP. Concurrently, EPC1 and EPC2 cells underwent autophagy following EtOH exposure, as evidenced by flow cytometry for Cyto-ID, which detects autophagic vesicles, and immunoblots demonstrating induction of the lipidated and cleaved form of LC3B and downregulation of SQSTM1/p62. In EPC1 and EPC2, pharmacological inhibition of autophagy flux by chloroquine increased mitochondrial oxidative stress while decreasing cell viability. In EPC2, autophagy induction was coupled with phosphorylation of AMP activated protein kinase (AMPK), a cellular energy sensor responding to low ATP levels, and dephosphorylation of downstream substrates of mechanistic Target of Rapamycin Complex (mTORC)-1 signaling. Pharmacological AMPK activation by AICAR decreased EtOH-induced reduction of ΔΨm and ATP in EPC2. Taken together, acute EtOH exposure leads to mitochondrial dysfunction and oxidative stress in esophageal keratinocytes, where the AMPK-mTORC1 axis may serve as a regulatory mechanism to activate autophagy to provide cytoprotection against EtOH-induced cell injury.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0239625PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7510980PMC
November 2020

Scale-Free Soft Sensor for Monitoring of Water Content in Fluid Bed Granulation Process.

Chem Pharm Bull (Tokyo) 2020 ;68(9):855-863

Department of Systems Science, Kyoto University.

In-line monitoring of granule water content during fluid bed granulation is important to control drug product qualities. In this study, a practical scale-free soft sensor to predict water content was proposed to cope with the manufacturing scale changes in drug product development. The proposed method exploits two key ideas to construct a scale-free soft sensor. First, to accommodate the changes in the manufacturing scale, the process parameters (PPs) that are critical to water content at different manufacturing scales were selected as input variables. Second, to construct an accurate statistical model, locally weighted partial least squares regression (LW-PLSR), which can cope with collinearity and nonlinearity, was utilized. The soft sensor was developed using both laboratory (approx. 4 kg) data and pilot (approx. 25 kg) scale data, and the prediction accuracy in the commercial (approx. 100 kg) scale was evaluated based on the assumption that the process was scaled-up from the pilot scale to the commercial scale. The developed soft sensor exhibited a high prediction accuracy, which was equivalent to the commonly used near-infrared (NIR) spectra-based method. The proposed method requires only standard instruments; therefore, it is expected to be a cost-effective alternative to the NIR spectra-based method.
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http://dx.doi.org/10.1248/cpb.c20-00315DOI Listing
April 2021

Coupling day length data and genomic prediction tools for predicting time-related traits under complex scenarios.

Sci Rep 2020 08 7;10(1):13382. Epub 2020 Aug 7.

Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo, 113-8657, Japan.

Genomic selection (GS) has proven to be an efficient tool for predicting crop-rank performance of untested genotypes; however, when the traits have intermediate optima (phenology stages), this implementation might not be the most convenient. GS might deliver high-rank correlations but incurring in serious bias. Days to heading (DTH) is a crucial development stage in rice for regional adaptability with a significant impact on yield potential. The objective of this research consisted in develop a novel method that accurately predicts time-related traits such as DTH in unobserved environments. For this, we propose an implementation that incorporates day length information (DL) in the prediction process for two relevant scenarios: CV0, predicting tested genotypes in unobserved environments (C method); and CV00, predicting untested genotypes in unobserved environments (CB method). The use of DL has advantages over weather data since it can be determined in advance just by knowing the location and planting date. The proposed methods showed that DL information significantly helps to improve the predictive ability of DTH in unobserved environments. Under CV0, the C method returned a root-mean-square error (RMSE) of 3.9 days, a Pearson correlation (PC) of 0.98 and the differences between the predicted and observed environmental means (EMD) ranged between -4.95 and 4.67 days. For CV00, the CB method returned an RMSE of 7.3 days, a PC of 0.93 and the EMD ranged between -6.4 and 4.1 days while the conventional GS implementation produced an RMSE of 18.1 days, a PC of 0.41 and the EMD ranged between -31.5 and 28.7 days.
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http://dx.doi.org/10.1038/s41598-020-70267-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7415153PMC
August 2020

Trialkylphosphines Having a Bulky Phosphacyclopentane Backbone: Structural and Redox Properties Depending on the Exocyclic Alkyl Groups and EPR Observation of a Persistent Trialkylphosphine Radical Cation.

J Org Chem 2020 Nov 7;85(22):14634-14642. Epub 2020 Aug 7.

Department of Chemistry, Graduate School of Science, Tohoku University, Aoba-ku, Sendai 980-8578, Japan.

Bulky phosphines and their redox properties have received increased attention in the view of useful auxiliary ligands for transition metal catalysts and Lewis-base components of frustrated Lewis pairs for chemical transformations. Herein we report the synthesis, structure, and properties of a series of trialkylphosphines (R = methyl, ethyl, isopropyl, -butyl, 1-adamantyl) that possess the bulky 2,2,5,5-tetrakis(trimethylsilyl)-1-phosphacyclopentane as a structural backbone. Among these phosphines, , which contains an adamantyl moiety, has a very large buried volume (%) for a trialkylphosphine (62.0) and shows a quasi-reversible oxidative wave at a lower oxidation potential (-0.12 V in CHCl, vs ferrocene/ferrocenium couple) by cyclic voltammetry. The reaction of with AgPF afforded a cationic silver aquo complex [Ag()(HO)][PF], whereas the reaction with NOSbF gave a persistent phosphine radical cation []. Based on the EPR spectra and DFT studies, the spin and positive charge of [] are localized on the phosphorus atom.
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http://dx.doi.org/10.1021/acs.joc.0c01393DOI Listing
November 2020

Olig2-Induced Semaphorin Expression Drives Corticospinal Axon Retraction After Spinal Cord Injury.

Cereb Cortex 2020 10;30(11):5702-5716

Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.

Axon regeneration is limited in the central nervous system, which hinders the reconstruction of functional circuits following spinal cord injury (SCI). Although various extrinsic molecules to repel axons following SCI have been identified, the role of semaphorins, a major class of axon guidance molecules, has not been thoroughly explored. Here we show that expression of semaphorins, including Sema5a and Sema6d, is elevated after SCI, and genetic deletion of either molecule or their receptors (neuropilin1 and plexinA1, respectively) suppresses axon retraction or dieback in injured corticospinal neurons. We further show that Olig2+ cells are essential for SCI-induced semaphorin expression, and that Olig2 binds to putative enhancer regions of the semaphorin genes. Finally, conditional deletion of Olig2 in the spinal cord reduces the expression of semaphorins, alleviating the axon retraction. These results demonstrate that semaphorins function as axon repellents following SCI, and reveal a novel transcriptional mechanism for controlling semaphorin levels around injured neurons to create zones hostile to axon regrowth.
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http://dx.doi.org/10.1093/cercor/bhaa142DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8179623PMC
October 2020