ABSTRACT
Vitamin D plays an important pleiotropic role in maintaining global homeostasis of the human body. Its functions go far beyond skeletal health, playing a crucial role in a plethora of cellular functions, as well as in extraskeletal health, ensuring the proper functioning of multiple human organs, including the skin. Genes from the Grainyhead-like (GRHL) family code for transcription factors necessary for the development and maintenance of various epithelia. Even though they are involved in many processes regulated by vitamin D, a direct link between vitamin D-mediated cellular pathways and GRHL genes has never been described. We employed various bioinformatic methods, quantitative real-time PCR, chromatin immunoprecipitation, reporter gene assays, and calcitriol treatments to investigate this issue. We report that the vitamin D receptor (VDR) binds to a regulatory region of the Grainyhead-like 1 (GRHL1) gene and regulates its expression. Ectopic expression of VDR and treatment with calcitriol alters the expression of the GRHL1 gene. The evidence presented here indicates a role of VDR in the regulation of expression of GRHL1 and correspondingly a role of GRHL1 in mediating the actions of vitamin D.
Subject(s)
Gene Expression Regulation , Receptors, Calcitriol , Transcription Factors , Receptors, Calcitriol/genetics , Receptors, Calcitriol/metabolism , Humans , Transcription Factors/metabolism , Transcription Factors/genetics , Gene Expression Regulation/drug effects , Calcitriol/pharmacology , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Protein Binding , Promoter Regions, Genetic , Repressor ProteinsABSTRACT
Ewing sarcoma is an aggressive malignancy of bone and soft tissue that accounts for â¼2% of cases of childhood cancer. It has been rarely reported as a secondary neoplasm. Data from the Childhood Cancer Survivor Study has evaluated secondary sarcomas in 5-year survivors of childhood cancer. We report 2 pediatric patients in northeast Pennsylvania, who developed secondary Ewing sarcoma of the 9th rib within 5 years of primary childhood leukemia diagnoses.
Subject(s)
Bone Neoplasms , Leukemia , Neoplasms, Second Primary , Sarcoma, Ewing , Sarcoma , Bone Neoplasms/therapy , Child , Humans , Ribs , Sarcoma, Ewing/therapyABSTRACT
A 15-year-old male presented with fatigue and weight loss for 1 month, petechiae and bruising for 2 weeks. He was diagnosed with concurrent new acute myeloid leukemia and coronavirus disease 2019. He was febrile and chest computed tomography scan showed ground glass opacities. Fever resolved after 4 days. Polymerase chain reaction test for coronavirus disease 2019 became negative after 2 days. Induction chemotherapy was initiated on day 8 and was complicated by multisystem inflammatory syndrome in children. The multisystem inflammatory syndrome in children was managed with symptomatic treatment and continued chemotherapy. Patient recovered and end of induction bone marrow showed remission of the leukemia.
Subject(s)
COVID-19/complications , Induction Chemotherapy/methods , Leukemia, Myeloid, Acute/complications , SARS-CoV-2/isolation & purification , Systemic Inflammatory Response Syndrome/pathology , Adolescent , COVID-19/pathology , COVID-19/virology , Humans , Leukemia, Myeloid, Acute/drug therapy , Leukemia, Myeloid, Acute/pathology , Leukemia, Myeloid, Acute/virology , Male , Remission Induction , Systemic Inflammatory Response Syndrome/drug therapy , Systemic Inflammatory Response Syndrome/virology , COVID-19 Drug TreatmentABSTRACT
Genes from the Grainyhead-like (GRHL) family code for transcription factors necessary for the development and maintenance of various epithelia. These genes are also very important in the development of many types of cancer. However, little is known about the regulation of expression of GRHL genes. Previously, there were no systematic analyses of the promoters of GRHL genes or transcription factors that bind to these promoters. Here we report that the Krüppel-like factor 4 (KLF4) and the paired box 5 factor (PAX5) bind to the regulatory regions of the GRHL genes and regulate their expression. Ectopic expression of KLF4 or PAX5 alters the expression of GRHL genes. In KLF4-overexpressing HEK293 cells, the expression of GRHL1 and GRHL3 genes was upregulated by 32% and 60%, respectively, whereas the mRNA level of GRHL2 gene was lowered by 28% when compared to the respective controls. The levels of GRHL1 and GRHL3 expression were decreased by 30% or 33% in PAX5-overexpressing HEK293 cells. The presence of minor frequency allele of single nucleotide polymorphism rs115898376 in the promoter of the GRHL1 gene affected the binding of KLF4 to this site. The evidence presented here suggests an important role of KLF4 and PAX5 in the regulation of expression of GRHL1-3 genes.
Subject(s)
Gene Expression Regulation , Kruppel-Like Transcription Factors/physiology , PAX5 Transcription Factor/physiology , Transcription Factors/metabolism , Animals , Chromatin Immunoprecipitation , Computer Simulation , DNA-Binding Proteins/metabolism , Electrophoretic Mobility Shift Assay , Gene Frequency , HEK293 Cells , Humans , Kruppel-Like Factor 4 , Kruppel-Like Transcription Factors/genetics , Mice , PAX5 Transcription Factor/metabolism , Polymorphism, Single Nucleotide/genetics , Real-Time Polymerase Chain Reaction , Repressor Proteins/metabolism , Transcription Factors/geneticsABSTRACT
Glanzmann Thrombasthenia is a rare bleeding disorder causing life-threatening bleeding at menarche in the adolescent female. Bleeding often necessitates admission and multiple blood transfusions. Due to the rarity of the disease, management of acute bleeding in new-onset menarche poses a particular challenge. A 12-year-old menarchial female had persistent vaginal bleeding despite multiple treatment modalities including aminocaproic acid, recombinant factor VIIa, intravenous estrogen, and gonadotropin receptor hormone agonists. Although the standard treatment of bleeding in patients with GT is primarily rFVIIa, new-onset menstrual bleeding in conjunction with an immature hypothalamic-pituitary-ovarian axis often requires expanding treatment to include multiple drug modalities. In our case, a two-step approach was necessary. The first is targeting the cessation of the first menses. The second is optimizing ongoing management of long-term control of heavy menstrual bleeding to achieve amenorrhea, prevent further hospital admissions, and avoid recurrent transfusions.
Subject(s)
DEAD-box RNA Helicases/genetics , Gene Deletion , Neoplastic Syndromes, Hereditary/diagnosis , Neoplastic Syndromes, Hereditary/genetics , Ribonuclease III/genetics , Adolescent , Alleles , Chromosomes, Human, Pair 14 , Comparative Genomic Hybridization , Female , Germ-Line Mutation , Heterozygote , Humans , Magnetic Resonance Imaging , Pedigree , PhenotypeABSTRACT
Primary leptomeningeal primitive neuroectodermal tumors (PNETs) are extremely rare childhood central nervous system malignancies harboring a very poor prognosis. There is no consensus treatment for these tumors to date. We report a case of a 10-year-old male who presented with mental status change, hydrocephalus, intracranial and spinal diffuse leptomeningeal enhancement without a primary mass upon cranial imaging and a negative initial biopsy until five months into his presentation. He responded significantly well to initial chemotherapy and radiotherapy.
Subject(s)
Meningeal Neoplasms/complications , Neuroectodermal Tumors, Primitive/complications , Biopsy , Child , Humans , Hydrocephalus/etiology , Male , Meningeal Neoplasms/drug therapy , Meningeal Neoplasms/radiotherapy , Neuroectodermal Tumors, Primitive/drug therapy , Neuroectodermal Tumors, Primitive/radiotherapy , PrognosisABSTRACT
Detour tasks are commonly used to study problem solving skills and inhibitory control in canids and primates. However, there is no comparable detour test designed for rodents despite its significance for studying the development of executive skills. Furthermore, mice offer research opportunities that are not currently possible to achieve when primates are used. Therefore, the aim of the study was to translate the classic detour task to mice and to compare obtained data with key findings obtained previously in other mammals. The experiment was performed with V-shaped barriers and was based on the water escape paradigm. The study showed that an apparently simple task requiring mice to move around a small barrier constituted in fact a challenge that was strongly affected by the visibility of the target. The most difficult task involved a completely transparent barrier, which forced the mice to resolve a conflict between vision and tactile perception. The performance depended both on the inhibitory skills and on previous experiences. Additionally, all mice displayed a preference for one side of the barrier and most of them relied on the egocentric strategy. Obtained results show for the first time that the behavior of mice subjected to the detour task is comparable to the behavior of other mammals tested previously with free-standing barriers. This detailed characterization of the detour behavior of mice constitutes the first step toward the substitution of rodents for primates in laboratory experiments employing the detour task.
Subject(s)
Behavior, Animal/physiology , Learning/physiology , Motor Activity/physiology , Spatial Behavior/physiology , Animals , Male , Mice , Problem Solving/physiologyABSTRACT
Abnormalities of coagulation have been reported in patients with thyroid dysfunction, although there is no clear information to explain the mechanism behind such irregularities. We are presenting a case of subclinical hypothyroidism that came with coagulation disturbance (hypocoagulation). Her symptoms resolved with leothyroxin in 4 weeks. Further studies are needed to clear the pathogenesis of the coagulation disturbance in such setting. The article also outlines some patents on hypothyroidism and hyperthyroidism.
Subject(s)
Blood Coagulation Disorders/physiopathology , Hypothyroidism/physiopathology , Adolescent , Blood Coagulation Disorders/etiology , Female , Hemostasis/physiology , Humans , Hypothyroidism/complications , Hypothyroidism/drug therapy , Thyroid Hormones/therapeutic use , Thyroxine/therapeutic useABSTRACT
Morphoproteomic analysis reveals the constitutive activation of the mTOR, ERK, and NF-kappaB pathways in high risk neuroblastoma (HRN) cases as evidenced by (a) collective commonalities of: phosphorylated (p)-mTOR, p70S6K, ERK 1/2, and NF-kappaBp65 protein analytes using phosphospecific probes directed against sites of activation; (b) nuclear translocation of p-p70S6K, p-ERK 1/2, and p-NF-kappaBp65; and (c) correlative expression of the S phase-associated kinase Skp-2 (at a relatively high percentage in tumoral nuclei) and of the anti-apoptotic protein bcl-2. Based on a review of the literature, these preliminary observations appear to be the first morphoproteomic study on primary neuroblastomas.
Subject(s)
Brain Neoplasms/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , NF-kappa B p50 Subunit/metabolism , Neuroblastoma/metabolism , Protein Kinases/metabolism , Proteomics , Antigens, Neoplasm/analysis , Antigens, Neoplasm/metabolism , Biomarkers, Tumor/analysis , Biomarkers, Tumor/metabolism , Brain Neoplasms/pathology , Cell Cycle/physiology , Cell Nucleus/metabolism , Cell Nucleus/pathology , Extracellular Signal-Regulated MAP Kinases/analysis , Humans , NF-kappa B p50 Subunit/analysis , Neoplasm Staging , Neuroblastoma/pathology , Protein Kinases/analysis , Ribosomal Protein S6 Kinases, 70-kDa/metabolism , S-Phase Kinase-Associated Proteins/metabolism , TOR Serine-Threonine KinasesABSTRACT
Alterations in apoptotic mechanisms favoring cell survival may be vital for modifying tumor behavior. Survivin, an inhibitor of apoptosis, and caspase 8, a proapoptotic enzyme, are key players in cellular apoptotic mechanisms. We investigated whether the levels of survivin and caspase 8 and the ratio between these 2 apoptotic factors correlate with tumor biology and predicts outcome in patients with neuroblastoma. Survivin and caspase 8 levels were quantified in 38 primary tumor specimens and analyzed individually and in relation to each other. High survivin expression and high survivin:caspase 8 ratios were associated with MYCN amplification, unfavorable histology, and high-risk group of disease (P<0.0008). High survivin mRNA levels were associated with worse overall survival (P=0.02) although the median follow up was only 22.6 months with a range of 1 day to 3.3 years. Low caspase 8 expression was associated with stage 4 disease, high-risk group, MYCN amplification, and unfavorable histology. Although the survivin:caspase 8 ratio was associated with these risk factors, the ratio did not improve the predictive value of survivin alone in this small series. Quantifying multiple apoptotic genes in neuroblastoma may supplement current risk stratification. Moreover, categorizing aberrant apoptotic gene expression in neuroblastoma may translate into novel therapeutic targets.
Subject(s)
Microtubule-Associated Proteins/genetics , Neoplasm Proteins/genetics , Neuroblastoma/genetics , RNA, Messenger/biosynthesis , Caspase 8 , Caspases/biosynthesis , Caspases/genetics , Cell Line, Tumor , Follow-Up Studies , Gene Amplification , Gene Expression Profiling , Humans , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins/biosynthesis , N-Myc Proto-Oncogene Protein , Neoplasm Proteins/biosynthesis , Neuroblastoma/metabolism , Neuroblastoma/therapy , Nuclear Proteins/biosynthesis , Nuclear Proteins/genetics , Oncogene Proteins/biosynthesis , Oncogene Proteins/genetics , Proportional Hazards Models , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , Risk Factors , Survival Rate , Survivin , Treatment OutcomeABSTRACT
PURPOSE: Novel treatment strategies for high-risk and disseminated neuroblastoma (NB) are actively sought because of the dismal prognosis of advanced stage disease. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a recently identified member of the tumor necrosis factor family. TRAIL is capable of inducing apoptosis in multiple tumor cell types, with little or no cytotoxicity against normal cells. EXPERIMENTAL DESIGN: We examined the activation and regulation of TRAIL-induced apoptosis in several human NB cell lines. The effect of TRAIL was examined in the context of TRAIL receptor (TRAIL-R) and survivin (an antiapoptotic protein) expression in the cell lines. The ratio of survivin/TRAIL-R messenger RNA was determined and evaluated as a marker of recurrent disease in patients with NB. RESULTS: TRAIL induced apoptotic cell death of NB with variable sensitivities among the cell lines tested. Compared with a sensitive cell line (early passage NB16), the resistant cell lines (NB7 and late passage NB16) expressed lesser amounts of the death-inducing TRAIL-R1 and R2, and greater levels of survivin, an inhibitor of apoptosis. TRAIL sensitivity was enhanced in resistant cell lines by treating with etoposide that concomitantly increased TRAIL-R expression and diminished survivin expression. Survivin overexpression in a TRAIL-sensitive NB line (early passage NB16) rendered it less sensitive to treatment with TRAIL. Conversely, inhibiting survivin expression in NB3 by antisense oligonucleotides enhanced TRAIL sensitivity. A high survivin/TRAIL-R ratio accurately predicted risk for recurrent disease in primary tumor specimens tested. CONCLUSIONS: These findings suggest that TRAIL therapy in combination with specific chemotherapeutic agents may represent an effective therapeutic strategy for NB. The cell's sensitivity to TRAIL is at least partially governed by both TRAIL-R and survivin expression, whereas the ratio between these 2 factors appears to have prognostic value in patients with this disease.
Subject(s)
Apoptosis Regulatory Proteins/pharmacology , Apoptosis/drug effects , Membrane Glycoproteins/pharmacology , Microtubule-Associated Proteins/pharmacology , Neoplasm Proteins/pharmacology , Neuroblastoma/metabolism , Receptors, Tumor Necrosis Factor/biosynthesis , Tumor Necrosis Factor-alpha/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Cell Line, Tumor , Child , Etoposide/pharmacology , GPI-Linked Proteins , Ganglioneuroblastoma/metabolism , Ganglioneuroma/metabolism , Humans , Inhibitor of Apoptosis Proteins , Neoplasm Recurrence, Local , Predictive Value of Tests , Receptors, TNF-Related Apoptosis-Inducing Ligand , Receptors, Tumor Necrosis Factor, Member 10c , Survivin , TNF-Related Apoptosis-Inducing Ligand , Tumor Necrosis Factor Decoy ReceptorsABSTRACT
Two patients with hemangioendothelioma and elevated plasma vascular endothelial growth factor (VEGF) levels are presented. A reduction in the plasma VEGF level after therapeutic intervention correlated with a successful clinical response. Conversely, a stable plasma VEGF level correlated with therapeutic failure.
Subject(s)
Biomarkers, Tumor/blood , Hemangioendothelioma/blood , Liver Neoplasms/blood , Neoplasm Proteins/blood , Soft Tissue Neoplasms/blood , Vascular Endothelial Growth Factor A/blood , Hemangioendothelioma/congenital , Hemangioendothelioma/surgery , Hepatectomy , Hip Contracture/etiology , Hip Contracture/pathology , Humans , Infant , Liver Neoplasms/congenital , Liver Neoplasms/surgery , Magnetic Resonance Imaging , Male , Neoplasm Recurrence, Local/drug therapy , Prednisolone/therapeutic use , Soft Tissue Neoplasms/drug therapy , Soft Tissue Neoplasms/surgery , Subcutaneous TissueABSTRACT
Important in the homeostasis of normal tissues, apoptosis can be altered to favor cell survival within tumors. High expression of survivin, an inhibitor of apoptosis, and absence of caspase 8, a pro-apoptotic enzyme, independently correlate with poor outcomes in several tumor types. Favorable histology Wilms tumor has a remarkably high cure rate; as a result, the focus of therapy is now aimed at reducing treatment-related morbidity. With the goal of safely reducing therapy in select subgroups of patients, the authors investigated whether the levels of apoptotic factors in tumors could predict the risk for recurrence. Tumor apoptotic factor levels were surveyed in a case-control study from the National Wilms Tumor Study 5 (NWTS-5) and measured via quantitative real-time RT-PCR. Survivin and caspase 8 levels were surveyed in 92 primary tumor specimens and SMAC, Bid, and CD95 were surveyed in 24 specimens. All four pro-apoptotic factors studied (caspase 8, SMAC, Bid, and CD95) were analyzed individually and in relation to survivin expression. Although survivin mRNA was present at markedly greater levels than in normal kidney, none of the factors assayed independently or as a ratio was associated with stage of disease or risk for tumor recurrence in this group of tumors.
Subject(s)
Apoptosis/physiology , Biomarkers, Tumor/analysis , Kidney Neoplasms/metabolism , Wilms Tumor/metabolism , BH3 Interacting Domain Death Agonist Protein , Carrier Proteins/metabolism , Case-Control Studies , Caspase 8 , Caspases/metabolism , Gene Expression , Inhibitor of Apoptosis Proteins , Kidney Neoplasms/pathology , Microtubule-Associated Proteins/metabolism , Mitochondrial Proteins/metabolism , Neoplasm Proteins , Prognosis , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction , Survivin , Wilms Tumor/pathology , fas Receptor/metabolismABSTRACT
BACKGROUND: Pediatric emergency RBC transfusions are often infused rapidly through 22-gauge (ga) or smaller needles or catheters using hand-held syringes. Data relating needle size, unit age, and infusion rate are needed to assess the risk of hemolysis and hyperkalemia in this setting. STUDY DESIGN AND METHODS: Multiple simulated transfusions were performed during storage of RBC units. Aliquots from five units were rapidly passed through needles (18, 20, 22-25 ga) using a hand-held syringe. Resulting plasma Hb and K+ concentrations were measured. Free Hb levels were used as a measure of needle-associated hemolysis (NAH). RESULTS: Passage through 18-ga and 20-ga needles caused no hemolysis, but rapid passage through 23-ga, 24-ga, and 25-ga did. RBCs stored less than 7 days showed significant K+ release with 23- to 25-ga needles. The greatest needle-associated K+ release was 10 mEq per L, on Day 5. Due to high K+ concentrations resulting from spontaneous efflux, K+ release from NAH was not detectable after 2 or more weeks of storage. CONCLUSIONS: Rapidly transfusing RBCs using hand-held syringes through 23-ga or smaller needles can cause hemolysis. In RBCs stored 2 weeks or more, NAH does not measurably increase K+ concentrations above that present from storage-related efflux. During rapid transfusions, RBC storage time is the primary risk factor for transfusion-associated hyperkalemia.
Subject(s)
Blood Transfusion/instrumentation , Hyperkalemia/etiology , Needles , Syringes , Blood Preservation , Hemoglobins , Hemolysis , Humans , Hyperkalemia/epidemiology , In Vitro Techniques , Potassium/metabolism , Risk FactorsABSTRACT
Granulocyte macrophage colony-stimulating factor (GM-CSF)-transduced autologous tumor cells form the basis of many immunotherapeutic strategies. We tested whether combining this approach with T-helper 1 (Th-1)-like immunostimulatory CpG oligodeoxynucleotides (CpG ODNs) would improve therapeutic efficacy in an established model of murine neuroblastoma. The weakly immunogenic Neuro-2a cell line was used in syngeneic A/J mice. CpG 1826 was tested for its antitumor effect alone and as an adjuvant to Neuro-2a cells retrovirally transduced to express murine GM-CSF (GM/Neuro-2a). Three days after wild-type (WT) tumor cell inoculation, mice in different groups were s.c. vaccinated in the opposite leg with combinations of WT neuro2a, irradiated (15 Gy) WT or GM/Neuro-2a transfectants with or without CpG 1826 (200 micro g). To test for the induction of memory responses, mice that rejected their tumor were rechallenged with WT Neuro-2a (1 x 10(6)) 7 weeks after vaccination. All of the mice in the control (unvaccinated) group died within 3 weeks after Neuro-2a inoculation. Most of the vaccinated groups had only minimal-to-modest antitumor responses, and the mice succumbed to tumor. Tumor growth was remarkably inhibited in the group of mice that received irradiated GM/Neuro-2a plus CpG and four (50%) of eight mice in this group survived tumor free. Tumor-free mice were resistant to further WT tumor cell challenge, indicating a memory response. Mechanistic studies showed that CpG alone induced a favorable Th-1-like cytokine immune response and vaccine-induced tumor cell killing was dependent on both CD4 and CD8 T cells that killed tumor cell targets by apoptosis. These results demonstrate that CpG ODNs enhanced the antitumor effect of irradiated GM-CSF secreting Neuro-2a cells. This vaccine strategy elicits a potent tumor antigen-specific immune response against established murine neuroblastoma and generates systemic neuroblastoma-specific immunity.