Cells Of Neural Crest Origin Research Articles

Autologous Transplantation of Skin-Derived Precursor Cells in a Porcine Model

BackgroundHirschprung's disease is characterized by aganglionic bowel and often requires surgical resection. Cell-based therapies have been investigated as potential alternatives to restore functioning neurons. Skin-derived precursor cells (SKPs) differentiate into neural and glial cells in vitro and generate ganglion-like structures in rodents. In this report, we aimed to translate this approach into a large animal model of aganglionosis using autologous transplantation of SKPs. MethodsJuvenile pigs underwent skin procurement from the shoulder and simultaneous chemical denervation of an isolated colonic segment. Skin cells were cultured in neuroglial-selective medium and labeled with fluorescent dye for later identification. The cultured SKPs were then injected into the aganglionic segments of colon, and the specimens were retrieved within seven days after transplantation. SKPs in vitro and in vivo were assessed with histologic samples for various immunofluorescent markers of multipotency and differentiation. SKPs from the time of harvest were compared to those at the time of injection using PCR. ResultsPrior to transplantation, 72% of SKPs stained positive for nestin and S100b, markers of neural and glial precursor cells of neural crest origin, respectively. Markers of differentiated neurons and gliocytes, TUJ1 and GFAP, were detected in 47% of cultured SKPs. After transplantation, SKPs were identified in both myenteric and submucosal plexuses of the treated colon. Nestin co-expression was detected in the SKPs within the aganglionic colon in vivo. Injected SKPs appeared to migrate and express early neuroglial differentiation markers. ConclusionsAutologous SKPs implanted into aganglionic bowel demonstrated immunophenotypes of neuroglial progenitors. Our results suggest that autologous SKPs may be potentially useful for cell-based therapy for patients with enteric nervous system disorders. Type of StudyBasic science.

A study of prognostic factors in cutaneous malignant melanoma

Background: Melanocytes are cells of neural crest origin that migrate principally to basement membrane of skin at dermo-epidermal junction. These cells when exposed to carcinogens result in malignant melanoma. Melanoma accounts for only 4% to 5% of all skin cancers but causes majority of deaths from skin cancers. Several factors affect its prognosis. Most reports agree that tumour thickness is the most important factor, but several others also interact.Methods: From the department of Medical College, Kottayam the cancer of patient who had histopathologically proven malignant melanoma during 2008-2012 were obtained from this HPR, details of the lesion such as Breslows thickness, level of invasion, presence of ulceration, mitoses, lymphocytic infiltration, tumor vascularity etc. were recorded. The case records of the some patients were then obtained from the record library, from which the clinical details of the lesions and follow up were recorded. These patients were followed till 2013. The significance and each of these factors were these analysed.Results: There were a total of 64 cases with equal gender distribution. The incidence reflected the global trend of increasing number of cases. The factors which affect prognosis included nodal status, thickness, level of invasion, ulceration, mitoses, lymphocyte infiltration, vascularity and lymphovascular emboli. A striking finding was most of the patients presented at an advanced stage, unlike the West.Conclusions: Malignant melanoma is a deadly disease especially when diagnosed late. Many factors in addition to thickness were found to have prognostic significance.

Runx2 function in cells of neural crest origin during intramembranous ossification

Runt-related transcription factor 2 (Runx2), also known as core binding factor 1 (Cbfa1), is a multifunctional transcription factor and an essential master gene controlling osteoblast differentiation. We previously demonstrated the in vivo functions of Runx2 in mesoderm-derived cells. However, no studies have been conducted on Runx2 function during the differentiation of neural crest (NC)-derived cells in vivo. Wingless-type MMTV integration site family member 1 (Wnt1) is expressed in the NC, and Wnt1-Cre efficiently targets craniofacial NC-derived cells. Runx2 deficiency in cells of the Wnt1 lineage (referred henceforth as Runx2wnt1−/− within mice) resulted in defective ossification in certain regions, primarily in the anterior half of the craniofacial bones, including the frontal bone, jugal bone, squamous temporal bone, mandible, maxilla, and nasal bone. The skeletal analysis also revealed that heterozygous Runx2wnt1+/− embryos had an impaired closure of the frontal bone at the metopic suture and lacked the secondary palate in spite of otherwise normal ossification. This result suggests that ossification at the central part of the frontal bone is more dependent on Runx2 expression in comparison to other areas. These results indicate that Runx2 is indispensable not only for mesoderm-derived cells but also for NC-derived cells to differentiate during intramembranous ossification after migration to their destination from the neural plate border. Moreover, this implies that there are different levels of dependency on Runx2 expression for successful ossification between NC-derived cells that have migrated to different locations.

Scaffold-Free Dental Pulp Cell Sheets to Enhance Facial Nerve Regeneration

Facial nerve defects may result from trauma, tumor ablation, and iatrogenic surgical injury. Facial paralysis has functional and aesthetic effects, including an asymmetric smile, oral incompetence, and lagophthalmos with a potential for corneal injury. These physical effects can have a significant psychosocial impact and result in a decline in the patient’s quality of life (1). Cable grafting comes with slow or incomplete recovery of volitional movement. In general, 6 to 12 months are required to see recovery of facial tone followed by onset of facial movement (2). Dental pulp cells (DPCs) are emerging as a promising source of stem cells in nerve regeneration mainly due to their neural crest cell origin. As derivatives of neural crest, DPCs produce neurotrophic factors (NTF) that may have a paracrine modulatory effect on damaged axons, ultimately enhancing axon regeneration and extension. DPCs are readily available from human exfoliated teeth. Our research group is investigating DPCs’ ability to form a scaffold-free cell sheet capable of producing NTFs that can be wrapped around facial nerve defects to accelerate facial nerve healing and enhance functional recovery.

Pediatric gastric ganglioneuroma presenting as anemia

Abstract Primary gastric masses are rare in childhood, and a gastric ganglioneuroma has not been reported in the pediatric population. In this report, we describe a 12-year-old female who presented with iron deficiency anemia and melena. Endoscopy was performed to elucidate the source of her symptoms, and revealed a gastric mass with overlying ulceration. Following resection and pathologic examination, the mass was diagnosed as a solitary polypoid ganglioneuroma. A solitary polypoid ganglioneuroma is an uncommon, benign tumor of neural crest cell origin. They are most often asymptomatic and found incidentally, but can present with rectal bleeding, obstruction, pain, and changes in bowel function. Complete resection is the therapy of choice to prevent progression of symptoms or rare transformation into a malignant neuroblastic tumor, like neuroblastoma. As of the patient's last post-operative appointment, she was healthy with resolution of her anemia.

Keratocytes are induced to produce collagen type II: A new strategy for in vivo corneal matrix regeneration

The stroma, the middle layer of the cornea, is a connective tissue making up most of the corneal thickness. The stromal extracellular matrix (ECM) consists of highly organised lamellae which are made up of tightly packed fibrils primarily composed of collagens type I and V. This layer is interspersed with keratocytes, mesenchymal cells of neural crest origin. We have previously shown that adult corneal keratocytes exhibit phenotypic plasticity and can be induced into a neuronal phenotype. In the current study we evaluated the potential of keratocytes to produce collagen type II via phenotypic reprogramming with exogenous chondrogenic factors. The cornea presents a challenge to tissue engineers owing to its high level of organisation and the phenotypic instability of keratocytes. Traditional approaches based on a scar model do not support the engineering of functional stromal tissue. Type II collagen is not found in the adult cornea but is reported to be expressed during corneal development, raising the possibility of using such an approach to regenerate the corneal ECM. Keratocytes in culture and within intact normal and diseased tissue were induced to produce collagen type II upon treatment with transforming growth factor Beta3 (TGFβ3) and dexamethasone. In vivo treatment of rat corneas also resulted in collagen type II deposition and a threefold increase in corneal hardness and elasticity. Furthermore, the treatment of corneas and subsequent deposition of collagen type II did not cause opacity, fibrosis or scarring. The induction of keratocytes with specific exogenous factors and resulting deposition of type II collagen in the stroma can potentially be controlled by withdrawal of the factors. This might be a promising new approach for in vivo corneal regeneration strategies aimed at increasing corneal integrity in diseases associated with weakened ectatic corneal tissue such as keratoconus.

Overexpression of PDGFRA cooperates with loss of NF1 and p53 to accelerate the molecular pathogenesis of malignant peripheral nerve sheath tumors

Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive, frequently metastatic sarcomas that are associated with neurofibromatosis type 1 (NF1), a prominent inherited genetic disease in humans. Although loss of the NF1 gene predisposes to MPNST induction, relatively long tumor latency in NF1 patients suggests that additional genetic or epigenetic abnormalities are needed for the development of these nerve sheath malignancies. To study the molecular pathways contributing to the formation of MPNSTs in NF1 patients, we used a zebrafish tumor model defined by nf1 loss in a p53-deficient background together with the overexpression of either wild-type or constitutively activated PDGFRA (platelet-derived growth factor receptor-α) under control of the sox10 neural crest-specific promoter. Here we demonstrate the accelerated onset and increased penetrance of MPNST formation in fish overexpressing both the wild-type and the mutant PDGFRA transgenes in cells of neural crest origin. Interestingly, overexpression of the wild-type PDGFRA was even more potent in promoting transformation than the mutant PDGFRA, which is important because ~78% of human MPNSTs have expression of wild-type PDGFRA, whereas only 5% harbor activating mutations of the gene encoding this receptor. Further analysis revealed the induction of cellular senescence in zebrafish embryos overexpressing mutant, but not wild-type, PDGFRA, suggesting a mechanism through which the oncogenic activity of the mutant receptor is tempered by the activation of premature cellular senescence in an NF1-deficient background. Taken together, our study suggests a model in which overexpression of wild-type PDGFRA associated with NF1 deficiency leads to aberrant activation of downstream RAS signaling and thus contributes importantly to MPNST development—a prediction supported by the ability of the kinase inhibitor sunitinib alone and in combination with the MEK inhibitor trametinib to retard MPNST progression in transgenic fish overexpressing the wild-type receptor.

The "neuro" of neuroblastoma: Neuroblastoma as a neurodevelopmental disorder.

Neuroblastoma is a childhood cancer derived from cells of neural crest origin. The hallmarks of its enigmatic character include its propensity for spontaneous regression under some circumstances and its association with paraneoplastic opsoclonus, myoclonus, and ataxia. The neurodevelopmental underpinnings of its origins may provide important clues for development of novel therapeutic and preventive agents for this frequently fatal malignancy and for the associated paraneoplastic syndromes. Ann Neurol 2016;80:13-23.

Frequency of varicella zoster virus DNA in human adrenal glands.

Varicella zoster virus (VZV) becomes latent in ganglionic neurons derived from neural crest cells. Because the adrenal gland also contains medullary chromaffin cells of neural crest origin, we examined human adrenal glands and medullary chromaffin cell tumors (pheochromocytomas) for VZV and herpes simplex virustype 1 (HSV-1). We found VZV, but not HSV-1, DNA in 4/63 (6%) normal adrenal glands. No VZV transcripts or antigens were detected in the 4 VZVDNA-positive samples. No VZV or HSV-1 DNA was found in 21 pheochromocytomas.

Validation of Housekeeping Genes to Study Human Gingival Stem Cells and Their In Vitro Osteogenic Differentiation Using Real-Time RT-qPCR.

Gingival stem cells (GSCs) are recently isolated multipotent cells. Their osteogenic capacity has been validated in vitro and may be transferred to human cell therapy for maxillary large bone defects, as they share a neural crest cell origin with jaw bone cells. RT-qPCR is a widely used technique to study gene expression and may help us to follow osteoblast differentiation of GSCs. For accurate results, the choice of reliable housekeeping genes (HKGs) is crucial. The aim of this study was to select the most reliable HKGs for GSCs study and their osteogenic differentiation (dGSCs). The analysis was performed with ten selected HKGs using four algorithms: ΔCt comparative method, GeNorm, BestKeeper, and NormFinder. This study demonstrated that three HKGs, SDHA, ACTB, and B2M, were the most stable to study GSC, whereas TBP, SDHA, and ALAS1 were the most reliable to study dGSCs. The comparison to stem cells of mesenchymal origin (ASCs) showed that SDHA/HPRT1 were the most appropriate for ASCs study. The choice of suitable HKGs for GSCs is important as it gave access to an accurate analysis of osteogenic differentiation. It will allow further study of this interesting stem cells source for future human therapy.

Robot-Assisted Resection of a Large Posterior Mediastinal Mass

Introduction: Ganglioneuromas are rare, typically benign, tumors that arise from tissues that have a neural crest cell origin. They typically occur in patients ranging in age from 10 to 40 years and are classically found in the adrenal glands. Ganglioneuromas are frequently asymptomatic and discovered incidentally while another condition is being investigated. The video demonstrates a difficult dissection that offers technical advantages over traditional thoracoscopy instruments by the articulating instruments of the robot with run time of 4 minutes 31 seconds. Background: An 18-year-old woman who was being followed for scoliosis was found to have a large left chest mass on a chest x-ray. A chest CT scan revealed a 10×7.5×6.5-cm mass in the left upper chest, consistent with a bronchogenic cyst. She had no reports of fever, shortness of breath, or chest pain, although she did endorse frequent feelings of left chest/shoulder tightness. In an attempt to avoid a large thoracotomy or sternotomy, we proceeded with a robot-assisted resection of the large mass. Methods: The patient was laid supine with the left chest slightly elevated and her left arm extended above her head. She was intubated with a Carlens tube for single lung ventilation. With the left lung deflated, an 8-mm robotic trocar was placed in the left midaxillary line (sixth intercostal space), an 8-mm robotic trocar was placed in left anterior axillary line (fifth intercostal space), and an 8-mm trocar was placed posterior to scapula (fourth intercostal space). An additional 5-mm step trocar was placed as an assistant port, lower left midchest in the posterior axillary line. Using hook cautery and graspers with bipolar cautery, the tumor was dissected free from the surrounding tissues and the blood vessels cauterized. The mass seemed to arise from the left sympathetic ganglia chain, which had to be sacrificed for tumor removal. The tumor extended into the apex of the left chest and into the lower part of the left neck just behind the head of the clavicle. The tumor was resected free and placed into an endocatch bag, where it was cut to facilitate its removal. The anterior axillary line trocar was removed and widened to ∼5 cm to allow specimen removal. A 28F chest tube was used because the larger tube was less likely to clog, and the chest hole was already large enough to accommodate it. The operative time was 5.5 hours, with 5 to 10 minutes of setup for the robot. Final pathology revealed a ganglioneuroma. Results: She was admitted to the step down unit postoperatively with patient-controlled anesthesia pain control. She was transitioned to oral pain medicine with a general diet on postoperative day 1. Her chest tube was removed and she was discharged on postoperative day 2. She was seen in clinic the following month with some left arm numbness and slight symptoms of Horner's syndrome (left eye ptosis and miosis when tired). Twelve months later, she reports being asymptomatic from her Horner's syndrome. Conclusions: Robot-assisted thoracic surgery provides great 3D visualization and articulating instruments that can be used to dissect large intrathoracic tumors and possibly avoid a large thoracotomy or sternotomy. No competing financial interests exist. Runtime of video: 4 mins 31 secs This work was presented as a poster at the 2014 IPEG conference in Scotland.

A Rare Case of Intrapulmonary Ewing Sarcoma Presenting with Left Atrial Tumor Thrombus

A previously healthy 24-year-old woman presented with shortness of breath. Chest radiograph (Fig. 1A) showed complete right upper and lower (RUL) lobe collapse with leftward mediastinal shift and abrupt cutoff of the right mainstem bronchus suggestive of an obstructing lesion. Chest computed tomography (CT) (Fig. 1B, C) revealed a 9-cm RUL mass (white arrowheads) with endobronchial invasion and suspected mediastinal involvement (black arrows). Pathology from bronchoscopic cytology (Fig. 2) yielded translocation-positive Ewing sarcoma. Positron emission tomography-CT showed hypermetabolic osseous foci compatible with metastases (Fig. 1D, E) and a hypermetabolic RUL mass extending toward the left atrium (LA) (Fig. 1F, G). An 18-month combination chemotherapy of cyclophosphamide–doxorubicin–vincristine/ifosfamide–etoposide was started to downsize the tumor before a possible surgical resection. To clarify possible tumor invasion into the heart, echocardiography was performed, identifying a 2.8 × 2.2 cm mass (Fig. 1H) entering the LA through the pulmonary vein. The differential diagnosis was thrombus versus invading tumor. For further clarification, cardiovascular magnetic resonance (CMR) imaging was performed, revealing a well-circumscribed heterogeneous RUL mass with areas of necrosis and hemorrhage (Fig. 1I). The mass invaded the right hilum, obstructed the right mainstem bronchus, and grew into the LA through the right superior pulmonary vein (Fig. 1I, black arrow). In addition, comparison between coronal positron emission tomography-CT (Fig. 1F) and CMR (Fig. 1I) revealed additional tissue apposed to the hypermetabolic tumor thrombus. Early postcontrast CMR confirmed the presence of a bland LA thrombus (Fig. 1J, black arrow), adherent to the enhancing tumor thrombus (white arrowhead). On the basis of the findings obtained by CMR, the patient was immediately placed on anticoagulation therapy.FIGURE 2Photomicrographs of the H&E-stained cell-block sample (upper panel, ×40 magnification) and Papanicolaou-stained bronchoscopic cytology sample (bottom panel, ×40 magnification) are shown. The H&E slide demonstrates a “small round blue cell” tumor with dark nuclei and minimal cytoplasm consistent with Ewing sarcoma cells. The Papanicolaou-stained slide shows better morphology and nuclear characteristics with central nucleoli and a rare mitosis. The characteristic cytologic finding in Ewing sarcomas is a dimorphic cell population; one being larger with a more pale nucleus (black arrow, pink/clear vacuoles in the pale cytoplasm) and the second with smaller size and darker nucleus (surrounding the pale cells).3Sanati S Lu DW Schmidt E Perry A Dehner LP Pfeifer JD Cytologic diagnosis of Ewing sarcoma/peripheral neuroectodermal tumor with paired prospective molecular genetic analysis.Cancer. 2007; 111: 192-199Crossref PubMed Scopus (43) Google Scholar H&E, hematoxilyn–eosin.View Large Image Figure ViewerDownload (PPT) The Ewing sarcoma family of tumors has a neural crest cell origin. Most of these tumors exhibit a translocation between chromosomes 11 and 22.1Grier HE The Ewing family of tumors. Ewing's sarcoma and primitive neuroectodermal tumors.Pediatr Clin North Am. 1997; 44: 991-1004Abstract Full Text Full Text PDF PubMed Scopus (259) Google Scholar The majority of tumors typically affect the skeleton. Rare manifestations, however, have been reported in several extraosseal tissues.1Grier HE The Ewing family of tumors. Ewing's sarcoma and primitive neuroectodermal tumors.Pediatr Clin North Am. 1997; 44: 991-1004Abstract Full Text Full Text PDF PubMed Scopus (259) Google Scholar Primary pulmonary Ewing tumor is an extremely rare manifestation of the extraosseus Ewing sarcomas (EES). As of today, the number of primary pulmonary EES cases reported in literature is less than 15, including only one case of primary pulmonary EES invading the heart.2Paul S Ramanathan T Cohen DM et al.Primary Ewing sarcoma invading the heart: resection and reconstruction.J Thorac Cardiovasc Surg. 2007; 133: 1667-1669Abstract Full Text Full Text PDF PubMed Scopus (10) Google Scholar Our case report highlights the importance of multimodality imaging for the diagnostic assessment of the primary tumor, tumor direct invasion, and metastasis and emphasizes the role of CMR in the differential diagnosis of intracardiac tumor mass and thrombus.

Patient-derived glioblastoma stem cells are killed by CD133-specific CAR T cells but induce the T cell aging marker CD57.

The AC133 epitope of CD133 is a cancer stem cell (CSC) marker for many tumor entities, including the highly malignant glioblastoma multiforme (GBM). We have developed an AC133-specific chimeric antigen receptor (CAR) and show that AC133-CAR T cells kill AC133+ GBM stem cells (GBM-SCs) both in vitro and in an orthotopic tumor model in vivo. Direct contact with patient-derived GBM-SCs caused rapid upregulation of CD57 on the CAR T cells, a molecule known to mark terminally or near-terminally differentiated T cells. However, other changes associated with terminal T cell differentiation could not be readily detected. CD57 is also expressed on tumor cells of neural crest origin and has been preferentially found on highly aggressive, undifferentiated, multipotent CSC-like cells. We found that CD57 was upregulated on activated T cells only upon contact with CD57+ patient-derived GBM-SCs, but not with conventional CD57-negative glioma lines. However, CD57 was not downregulated on the GBM-SCs upon their differentiation, indicating that this molecule is not a bona fide CSC marker for GBM. Differentiated GBM cells still induced CD57 on CAR T cells and other activated T cells. Therefore, CD57 can apparently be upregulated on activated human T cells by mere contact with CD57+ target cells.

Echocardiographic evidence of innate aortopathy in the human intracranial aneurysm.

BackgroundIntracranial aneurysm (IA) is significantly more prevalent in patients with coarctation of the aorta or bicuspid aortic valve than in the general population, suggesting a common pathophysiology connecting IA and aortopathy. Here, we analyzed echocardiographic aortic root dimension (ARD) in patients with IA to confirm this possibility.MethodsFrom January 2008 to December 2010, 260 consecutive patients with IA who were admitted to our institution for coil embolization or for acute stroke management and who also underwent echocardiography were enrolled. We hypothesized that patients with large, ruptured, or multiple IAs are more likely to harbor co-prevalent aortopathy as measured by ARD compared to patients with small, isolated, unruptured IAs. Eccentric group was defined as patients aged <55 years with at least one ruptured aneurysm, an aneurysm ≥7 mm in size, or multiple aneurysms; the remainder was classified into a non-eccentric group. Clinical, angiographic, and echocardiographic findings of the two groups were compared.ResultsARD was significantly larger in the eccentric group than in the non-eccentric group (P = 0.049), and the difference was confirmed by multivariable analysis (P = 0.02). Subgroup analysis of patients aged <55 years showed similar result for ARD (P = 0.03), whereas hypertension was more associated with the non-eccentric group (P = 0.01). In addition, height was inversely related to aneurysm size after adjustment for age, sex, weight, ARD, smoking status, and number of aneurysms (P = 0.004).ConclusionsA certain group of IA patients share a common intrinsic wall defect with aortopathy. Shared neural crest cell origin may give rise to this phenomenon.

Formation of Persistent Hyperplastic Primary Vitreous in Ephrin-A5−/−Mice

Primary vitreous regression is a critical event in mammalian eye development required for proper ocular maturity and unhindered vision. Failure of this event results in the eye disease persistent hyperplastic primary vitreous (PHPV), also identified as persistent fetal vasculature (PFV), a condition characterized by the presence of a fibrovascular mass adjacent to the lens and retina, and associated with visual disability and blindness. Here, we identify ephrin-A5 to be a critical regulator for primary vitreous regression. Wild-type and ephrin-A5(-/-) eyes were examined at various developmental stages to determine the progression of PHPV. Eye tissue was sectioned and examined by H&E staining. Protein expression and localization was determined through immunohistochemistry. Relative levels of Eph receptors were determined by RT-PCR. Ephrin-A5(-/-) animals develop ocular phenotypes representative of PHPV, most notably the presence of a large hyperplastic mass posterior to the lens that remains throughout the lifetime of the animal. The aberrant tissue in these mutant mice consists of residual hyaloid vessels surrounded by pigmented cells of neural crest origin. Labeling with bromodeoxyuridine (BrdU) and detection of proliferating cell nuclear antigen (PCNA) expression shows that the mass in ephrin-A5(-/-) animals is mitotically active in embryonic and postnatal stages. Ephrin-A5 is a critical factor that regulates primary vitreous regression.

Malignant peripheral nerve sheath tumour of penis

Malignant peripheral nerve sheath tumour (MPNST) is a rare variety of soft tissue sarcoma that originates from Schwann cells or pluripotent cells of neural crest origin. They have historically been difficult tumours to diagnose and treat. Surgery is the mainstay of treatment with a goal to achieve negative margins. Despite aggressive surgery and adjuvant therapy, the prognosis of patients with MPNST remains poor. MPNST arising from penis is a very rare entity; thus, it presents a diagnostic and therapeutic challenge. We present a case of penile MPNST in a 38-year-old man in the absence of neurofibromatosis treated with surgery followed by post-operative radiotherapy to a dose of 60 Gray in 30 fractions and adjuvant chemotherapy with ifosfamide and adriamycin.

Congenital central hypoventilation syndrome and intestinal aganglionosis: A case report

Congenital central hypoventilation syndrome (CCHS), also called ‘Ondine’s curse’, is characterised by an abnormal ventilatory response to progressive hypercapnia and sustained hypoxaemia. Neonates with this condition experience hypoventilation or apnoea while asleep. Patients may also have congenital intestinal aganglionosis (CIA), aganglionic megacolon or Hirschsprung’s disease, suggesting an aberrant phenotype arising from a defect of migration or diff erentiation of neural crest cells. Some patients also have tumours of neural crest cell origin, including neuroblastoma, ganglioneuroma and ganglioneuroblastoma. Th e association of CCHS and CIA is called Ondine- Hirschsprung disease (Haddad syndrome). A few cases have been diagnosed in South Africa, but none has been reported. We report a case of CCHS and CIA with a PHOX2B gene mutation.

Juxta-adrenal malignant schwannoma with lymph node metastases

Juxta-adrenal malignant schwannoma is a very rare tumour of neural crest cell origin. A 69-year-old woman visited for proper management of incidentally detected adrenal mass as part of a dyspepsia workup. Metabolic evaluation was unremarkable and imaging results suggested a malignant adrenal mass with lymph node metastases. The patient underwent a left adrenalectomy with para-aortic lymphadenectomy. The diagnosis and its origin was confirmed by histologic and immunohistochemical studies. We report the first case of juxta-adrenal malignant schwannoma with lymph node metastases, and review the literature on this rare tumour.

Functional Corneal Endothelium Derived from Corneal Stroma Stem Cells of Neural Crest Origin by Retinoic Acid and Wnt/β-Catenin Signaling

Corneal endothelial dysfunction remains a major indication for corneal transplantation. Both corneal endothelial cells and stromal cells originate from the neural crest, but have distinct phenotypes and function in the adult cornea. We previously reported that stem cells isolated from the adult corneal stroma [cornea-derived precursors (COPs)] show characteristics of multipotent neural crest-derived stem cells. In this study, we report the induction of functional tissue-engineered corneal endothelium (TECE) from mouse and human COPs. TECE was engineered from Wnt1-Cre/Floxed EGFP mouse COPs in a medium containing retinoic acid and glycogen synthase kinase (GSK) 3β inhibitor (activator of Wnt/β-catenin signaling). The expression levels of major markers characterizing corneal endothelial function (Atp1a1, Slc4a4, Car2, Col4a2, Col8a2, and Cdh2) were significantly upregulated. Both retinoic acid and GSK 3β inhibitor upregulated the expression of Pitx2, a homeobox gene involved in the development of the anterior segment of the eye. GSK 3β inhibitor increased Atp1a1 expression and Na,K-ATPase pump activity of TECE, which was significantly higher than COPs or control 3T3 cells, and 2.6-fold higher than cultured mouse corneal endothelial cells. Mouse TECE transplanted into rabbit corneas maintained transparency and corneal thickness, whereas control corneas without TECE showed marked edema and increased corneal thickness. Furthermore, we successfully induced TECE from human COPs, and human TECE transplanted into rabbit corneas also maintained corneal transparency and thickness. This protocol enables efficient production of corneal endothelium from corneal stromal stem cells by direct induction, which may lead to a novel stem cell therapy for corneal endothelial dysfunction.

Anterior mediastinal schwannoma leading to dyspnea: a case report

Schwannomas are rare tumors of neural crest cell origin. Most schwannomas occur in the head, neck and limbs. Thoracic schwannomas are extremely rare and mostly appear in the posterior mediastinum. There are few case reports of anterior mediastinal schwannoma. In this article, we report an anterior mediastinal schwannoma resected using median sternotomy in an 29-years-old female who presented with the complaint of dyspnea. In this article, we aimed to review the literature and discuss the anterior mediastinal localization of schwannoma.