The Oncolytic Activity of Zika Viral Therapy in Human Neuroblastoma In Vivo Models Confers a Major Survival Advantage in a CD24-dependent Manner.

Neuroblastoma is the most common extracranial tumor, accounting for 15% of all childhood cancer-related deaths. The long-term survival of patients with high-risk tumors is less than 40%, and MYCN amplification is one of the most common indicators of poor outcomes. Zika virus (ZIKV) is a mosquito-borne flavivirus associated with mild constitutional symptoms outside the fetal period. Our published data showed that high-risk and recurrent neuroblastoma cells are permissive to ZIKV infection, resulting in cell type-specific lysis. In this study, we assessed the efficacy of ZIKV as an oncolytic treatment for high-risk neuroblastoma using in vivo tumor models. Utilizing both MYCN-amplified and non-amplified models, we demonstrated that the application of ZIKV had a rapid tumoricidal effect. This led to a nearly total loss of the tumor mass without evidence of recurrence, offering a robust survival advantage to the host. Detection of the viral NS1 protein within the tumors confirmed that a permissive infection preceded tissue necrosis. Despite robust titers within the tumor, viral shedding to the host was poor and diminished rapidly, correlating with no detectable side effects to the murine host. Assessments from both primary pretreatment and recurrent posttreatment isolates confirmed that permissive sensitivity to ZIKV killing was dependent on the expression of CD24, which was highly expressed in neuroblastomas and conferred a proliferative advantage to tumor growth. Exploiting this viral sensitivity to CD24 offers the possibility of its use as a prognostic target for a broad population of expressing cancers, many of which have shown resistance to current clinical therapies. SIGNIFICANCE: Sensitivity to the tumoricidal effect of ZIKV on high-risk neuroblastoma tumors is dependent on CD24 expression, offering a prognostic marker for this oncolytic therapy in an extensive array of CD24-expressing cancers.

Concurrent dermoid and epidermoid cysts in an adolescent patient: a case report.

Dermoid and epidermoid cysts are benign lesions of ectodermal origin which are pathologically distinct entities, although often clinically indistinguishable. Cyst location, mobility, and appearance on MRI can help distinguish the two, however the distinction is mostly academic since both types have similar management. Co-occurrence of dermoid and epidermoid cysts together in the same patient has not been observed in the literature, however one case of an epidermoid cyst evolving into a dermoid cyst has been documented. In this case report, we identify a 16-year-old male with three separate cysts of the scalp and leg which, after histopathological analysis following surgical resection, were found to represent both dermoid and epidermoid cysts. We offer potential explanations for this rare occurrence in the absence of a genetic syndrome and highlight the importance of performing a thorough work-up of patients with multiple cysts.

Acute Case of Trichobezoar Diagnosed From Computed Tomography and 3D Images: Rapunzel Syndrome Re-examined.

A trichobezoar is a rare cause of abdominal pain due to an indigestible mass in the gastrointestinal tract that is composed of a patient's hair. If a trichobezoar grows and extends from the gastric body to the pylorus and into the small bowel, it is considered Rapunzel syndrome. We present a case of an 11-year-old female patient with Rapunzel syndrome who presented with four weeks of colicky abdominal pain, vomiting, constipation, and severe malnutrition. Computed tomography of the abdomen and pelvis with 3D rendering demonstrated a large bezoar, and the patient was successfully treated with exploratory laparotomy, gastrostomy, and removal of the trichobezoar intact.

Bowel Obstruction Secondary to Spontaneous Knot Formation of Ventriculoperitoneal Shunt.

Ventriculoperitoneal (VP) shunts are frequently placed for the treatment of hydrocephalus. Shunt complications are a common occurrence typically involving infection, disconnections, or blockages. Abdominal complications involving the intraperitoneal portion of the catheter are rare. Spontaneous peritoneal knot formation involving the bowel with subsequent obstruction is even rarer. Spontaneous knot formation of a VP shunt is also not commonly seen in the adult population. In this report, we present the case of an 18-year-old male with cerebral palsy and hydrocephalus requiring VP shunt placement who developed a spontaneous knot leading to bowel obstruction requiring emergency laparoscopic surgery.

Chaperonin containing TCP-1 (CCT/TRiC) is a novel therapeutic and diagnostic target for neuroblastoma.

Chaperonin containing TCP1 (CCT/TRiC) is a multi-subunit protein folding complex that enables the cancer phenotype to emerge from the mutational landscape that drives oncogenesis. We and others linked increased expression of CCT subunits to advanced tumor stage and invasiveness that inversely correlates with cancer patient outcomes. In this study, we examined the expression of the second CCT subunit, CCT2, using genomic databases of adult and pediatric tumors and normal tissues, and found that it was highly expressed in pediatric cancers, showing a significant difference compared to normal tissues. Histologic staining confirmed that CCT subunits are highly expressed in tumor tissues, which was exemplified in neuroblastoma. Using two neuroblastoma cells, MYCN-amplified, IMR-32 cells, and non-amplified, SK-N-AS cells, we assessed baseline levels for CCT subunits and found expressions comparable to the highly invasive triple-negative breast cancer (TNBC) cell line, MDA-MB-231. Exogenous expression of CCT2 in both SK-N-AS and IMR-32 cells resulted in morphological changes, such as larger cell size and increased adherence, with significant increases in the CCT substrates, actin, and tubulin, as well as increased migration. Depletion of CCT2 reversed these effects and reduced cell viability. We evaluated CCT as a therapeutic target in IMR-32 cells by testing a novel peptide CCT inhibitor, CT20p. Treatment with CT20p induced cell death in these neuroblastoma cells. The use of CCT2 as a biological indicator for detection of neuroblastoma cells shed in blood was examined by spiking IMR-32 cells into human blood and using an anti-CCT2 antibody for the identification of spiked cancer cells with the CellSearch system. Results showed that using CCT2 for the detection of neuroblastoma cells in blood was more effective than the conventional approach of using epithelial markers like cytokeratins. CCT2 plays an essential role in promoting the invasive capacity of neuroblastoma cells and thus offers the potential to act as a molecular target in the development of novel therapeutics and diagnostics for pediatric cancers.

Thyroid Teratoma in a Pediatric Patient.

Congenital thyroid teratomas are rare in the pediatric population as well as in the adult population. While they are typically found in the gonadal regions, extragonadally, they are commonly found in the sacrococcygeal region, with teratomas of the head and neck rarely found, comprising only about 1%-6% of all pediatric teratomas. Due to a concern for potential airway compromise and increased risk of malignancy with age, early surgical excision is recommended. In this case report, we present a two-year-old female who underwent laryngoscopy with subsequent right thyroid lobectomy for a large thyroid mass, which was found to be a congenital thyroid teratoma.

CD24 Expression Dampens the Basal Antiviral State in Human Neuroblastoma Cells and Enhances Permissivity to Zika Virus Infection.

Zika virus (ZIKV) exhibits distinct selectivity for infection of various cells and tissues, but how host cellular factors modulate varying permissivity remains largely unknown. Previous studies showed that the neuroblastoma cell line SK-N-AS (expressing low levels of cellular protein CD24) was highly restricted for ZIKV infection, and that this restriction was relieved by ectopic expression of CD24. We tested the hypothesis that CD24 expression allowed ZIKV replication by suppression of the antiviral response. SK-N-AS cells expressing an empty vector (termed CD24-low cells) showed elevated basal levels of phosphorylated STAT1, IRF-1, IKKE, and NFκB. In response to exogenously added type I interferon (IFN-I), CD24-low cells had higher-level induction of antiviral genes and activity against two IFN-I-sensitive viruses (VSV and PIV5-P/V) compared to SK-N-AS cells with ectopic CD24 expression (termed CD24-high cells). Media-transfer experiments showed that the inherent antiviral state of CD24-low cells was not dependent on a secreted factor such as IFN-I. Transcriptomics analysis revealed that CD24 expression decreased expression of genes involved in intracellular antiviral pathways, including IFN-I, NFκB, and Ras. Our findings that CD24 expression in neuroblastoma cells represses intracellular antiviral pathways support the proposal that CD24 may represent a novel biomarker in cancer cells for susceptibility to oncolytic viruses.

Delayed Presentation of Duodenal Atresia in a Male With Trisomy 21.

The duodenum is the secondmost common site of congenital intestinal obstruction. There are three types of congenital duodenal atresia according to the severity of obstruction. Duodenal atresia is thought to develop due to the failure of recanalization of the gut lumen during embryonic development. This congenital abnormality usually presents in utero or shortly after birth with signs of intestinal obstruction. However, rare cases can present later in life. In this case report, we will discuss a two-year-old male with trisomy 21 who presented with intractable vomiting and failure-to-thrive. He did not have the classic clinical or diagnostic signs of duodenal atresia, but on exploratory laparotomy, he was found to have severe duodenal stenosis. Diamond-shaped duodenoduodenostomy was performed to bypass the stenosed intestine. The patient recovered well from surgery and was able to tolerate a soft mechanical diet without vomiting one week postoperatively. This case exhibits a particularly delayed and atypical presentation of duodenal stenosis. Yet, it is imperative to recognize this presentation from an educational and clinical standpoint for surgical intervention.

Juvenile Retention Polyp in a Teenager.

The proper management of a prolapsed rectal mass in a child or teenager is challenging. Given that the underlying etiology of a prolapsed rectal mass in this population is not always immediately clear, interdisciplinary assessment is often required. Juvenile polyps, more commonly presenting with bleeding than a prolapsed mass, can mimic the appearance of both hemorrhoids and the rectum itself - making a purely clinical diagnosis difficult. Presented here is a case of a prolapsed colorectal polyp in a teenage boy, who underwent manual reduction of the mass, followed by colonoscopy and endoscopic ligation. Further histological evaluation revealed it to be a juvenile retention polyp. Despite the rarity of polyp prolapse as a presenting symptom, this case underscores the importance of considering colonic polyps as the etiology of a prolapsed anorectal mass in a teenager.

Submandibular Gland Injury With a Ball Bearing Gunshot Wound.

Submandibular gland injury is a rare occurrence that has been only documented in case reports. This is due to its protected location under the mandible, and only penetrating injuries to the floor of the mouth or trauma underneath the mandible can reach and damage it. While pediatric injuries due to non-powder firearms are decreasing yearly, 80.8% of the injuries were due to ball bearing (BB) guns. This case report explores the diagnosis and management of a 16-year-old girl who presented with a BB gunshot wound to the submandibular gland. The anatomy, imaging, and surgical management are detailed, and diagnosis guidelines and treatment options are analyzed and explained. This case highlights the importance of understanding the harm that non-powder firearms are capable of causing despite being perceived as toys.

Laparoscopic Resection of a Gastric Diverticulum in an Adolescent.

Gastric diverticula rarely occur in adolescence. In adults, they are predominantly congenital, asymptomatic, and are located adjacent to the gastroesophageal junction on the posterior aspect of the stomach wall. In this report we present a 14-year-old female who underwent laparoscopic gastric diverticulectomy after incidental discovery on magnetic resonance urography.

Keystone Flap Reconstruction after Resection of a Large Paraspinal Venous Malformation in an Infant.

The keystone flap is well known to plastic surgeons and is frequently utilized for its ease of implementation, limited donor site morbidity, and favorable aesthetic outcomes. Although keystone flaps have been described in reconstruction of myelomeningocele defects, there have been no reports of their application to infants with large vascular malformations. This case illustrates the utilization of a keystone flap in reconstruction of a large posterior trunk defect that resulted from excision of a massive venous malformation in an 8-week-old infant with blue rubber bleb nevus syndrome. The patient's consumptive coagulopathy resolved in the early postoperative period, and long-term follow-up demonstrated a favorable aesthetic outcome. This case reiterates the power and versatility of the keystone flap technique through its novel application to an infant with a life-threatening venous malformation on the posterior trunk.

Adolescent With VACTERL Association Presents With Recurrent Pneumonia.

VACTERL is a condition that includes various anatomic anomalies including vertebral, cardiac, tracheoesophageal fistula (TEF), renal, and limb defects. TEF can be found in up to 80% of patients with the condition. Although TEFs are usually identified early in life, the H-type TEF can be more difficult to detect. We report the case of a 15-year-old male with a previous diagnosis of VACTERL who presented with a history of recurrent pneumonia, chest pain, and asthma and was found to have a previously undetected H-type TEF that was surgically repaired. When evaluating a patient with features of VACTERL, it is important to choose studies that can explore the presence of all associated features. Clinical history and type of imaging utilized can be essential in making a timely diagnosis, especially for H-type TEF.

Laparoscopic Repair of Bilateral Congenital Diaphragmatic Hernia Without Pulmonary Hypoplasia.

We report the case of a 14-month-old female who had a right-sided congenital diaphragmatic hernia (CDH) without pulmonary hypoplasia. The patient was preoperatively diagnosed with a Morgagni hernia due to the size and location of the hernia seen on imaging. However, the patient was found to have bilateral diaphragmatic defects intraoperatively, and her right diaphragm was almost completely absent. Our patient did not have pulmonary hypoplasia or any of the respiratory comorbidities that CDH patients typically present with, though she did have repeated respiratory infections and cough. This case demonstrates that CDH is not always diagnosed in an accurate or timely manner radiographically and that the surgeon should be prepared to potentially repair more of the diaphragm than expected. Additionally, there is a need to study the pathophysiology and genetics of CDHs further.

The Killing of Human Neuroblastoma Cells by the Small Molecule JQ1 Occurs in a p53-Dependent Manner.

BACKGROUND: MYCN amplification is a prognostic biomarker associated with poor prognosis of neuroblastoma in children. The overall survival of children with MYCN-amplified neuroblastoma has only marginally improved within the last 20 years. The Bromodomain and Extra-Terminal motif (BET) inhibitor, JQ1, has been shown to downregulate MYCN in neuroblastoma cells. OBJECTIVE: To determine if JQ1 downregulation of MYCN in neuroblastomas can offer a target- specific therapy for this, difficult to treat, pediatric cancer. METHODS: Since MYCN-amplified neuroblastoma accounts for as much as 40 to 50 percent of all high-risk cases, we compared the effect of JQ1 on both MYCN-amplified and non-MYCN-amplified neuroblastoma cell lines and investigated its mechanism of action. RESULTS: In this study, we show that JQ1 can specifically target MYCN for downregulation, though this effect is not specific to only MYCN-amplified cells. And although we can confirm that the loss of MYCN alone can induce apoptosis, the exogenous rescue of MYCN expression can abrogate much of this cytotoxicity. More fascinating, however, was the discovery that the JQ1-induced knockdown of MYCN, which led to the loss of the human double minute 2 homolog (HDM2) protein, also led to the accumulation of tumor protein 53 (also known as TP53 or p53), which ultimately induced apoptosis. Likewise, the knockdown of p53 also blunted the cytotoxic effects of JQ1. CONCLUSION: These data suggest a mechanism of action for JQ1 cytotoxicity in neuroblastomas and offer a possible prognostic target for determining its efficacy as a therapeutic.

Zika virus as an oncolytic treatment of human neuroblastoma cells requires CD24.

Neuroblastoma is the second most common childhood tumor. Survival is poor even with intensive therapy. In a search for therapies to neuroblastoma, we assessed the oncolytic potential of Zika virus. Zika virus is an emerging mosquito-borne pathogen unique among flaviviruses because of its association with congenital defects. Recent studies have shown that neuronal progenitor cells are likely the human target of Zika virus. Neuroblastoma has been shown to be responsive to infection. In this study, we show that neuroblastoma cells are widely permissive to Zika infection, revealing extensive cytopathic effects (CPE) and producing high titers of virus. However, a single cell line appeared poorly responsive to infection, producing undetectable levels of non-structural protein 1 (NS1), limited CPE, and low virus titers. A comparison of these poorly permissive cells to highly permissive neuroblastoma cells revealed a dramatic loss in the expression of the cell surface glycoprotein CD24 in poorly permissive cells. Complementation of CD24 expression in these cells led to the production of detectable levels of NS1 expression after infection with Zika, as well as dramatic increases in viral titers and CPE. Complementary studies using the Zika virus index strain and a north African isolate confirmed these phenotypes. These results suggest a possible role for CD24 in host cell specificity by Zika virus and offer a potential therapeutic target for its treatment. In addition, Zika viral therapy can serve as an adjunctive treatment for neuroblastoma by targeting tumor cells that can lead to recurrent disease and treatment failure.

Nanoparticle delivery of curcumin induces cellular hypoxia and ROS-mediated apoptosis via modulation of Bcl-2/Bax in human neuroblastoma.

In this study, several formulations of nanoceria and dextran-nanoceria with curcumin, each demonstrated to have anti-cancer properties, were synthesized and applied as treatment for human childhood neuroblastoma. The anti-cancer activities of these formulations were explored in neuroblastoma models of both MYCN-amplified and non-amplified cell lines. Ceria nanoparticles, coated with dextran and loaded with curcumin, were found to induce substantial cell death in neuroblastoma cells (up to a 2-fold and a 1.6-fold decrease in cell viability for MYCN-upregulated and normal expressing cell lines, respectively; *p < 0.05) while producing no or only minor toxicity in healthy cells (no toxicity at 100 µM; **p < 0.01). This formulation evokes prolonged oxidative stress, stabilizing HIF-1α, and inducing caspase-dependent apoptosis (up to a 2.4-fold increase over control; *p < 0.05). Overall, nano-therapeutic treatments showed a more pronounced effect in MYCN-amplified cells, which are traditionally more resistant to drug therapies. These results represent a very promising alternative to small molecule drug therapies for robust cancers.

The long non-coding RNA GAS5 differentially regulates cell cycle arrest and apoptosis through activation of BRCA1 and p53 in human neuroblastoma.

The long non-coding RNA GAS5 has been shown to modulate cancer proliferation in numerous human cancer systems and has been correlated with successful patient outcome. Our examination of GAS5 in neuroblastoma has revealed robust expression in both MYCN-amplified and non-amplified cell lines. Knockdown of GAS5 In vitro resulted in defects in cell proliferation, apoptosis, and induced cell cycle arrest. Further analysis of GAS5 clones revealed multiple novel splice variants, two of which inversely modulated with MYCN status. Complementation studies of the variants post-knockdown of GAS5 indicated alternate phenotypes, with one variant (FL) considerably enhancing cell proliferation by rescuing cell cycle arrest and the other (C2) driving apoptosis, suggesting a unique role for each in neuroblastoma cancer physiology. Global sequencing and ELISA arrays revealed that the loss of GAS5 induced p53, BRCA1, and GADD45A, which appeared to modulate cell cycle arrest in concert. Complementation with only the FL GAS5 clone could rescue cell cycle arrest, stabilizing HDM2, and leading to the loss of p53. Together, these data offer novel therapeutic targets in the form of lncRNA splice variants for separate challenges against cancer growth and cell death.

Gastric metastasis from an alveolar soft part sarcoma in a child: case report and review of the literature.

The authors report a child with alveolar soft part sarcoma who developed significant anemia due to gastrointestinal blood loss. Evaluation revealed the source of bleeding as a gastric metastasis, which was successfully removed. A brief review of gastrointestinal involvement by alveolar soft part sarcoma is discussed.