GATA3 Research Articles

Hypoparathyroidism, deafness and renal dysplasia syndrome caused by a GATA3 splice site mutation leading to the activation of a cryptic splice site.

The HDR syndrome is a rare autosomal dominant disorder characterised by Hypoparathyroidism, Deafness, and Renal dysplasia, and is caused by inactivating heterozygous germline mutations in the GATA3 gene. We report an 11-year-old girl with HDR syndrome caused by a heterozygous mutation located at the splice acceptor site of exon 5 of the GATA3 gene (NM_001002295.2: c.925-1G>T). Functional studies using a minigene assay showed that this splice site mutation abolished the normal splicing of the GATA3 pre-mRNA and led to the use of a cryptic splice acceptor site, resulting in the loss of the first seven nucleotides (TCTGCAG) of exon 5 in the GATA3 mRNA. These findings increase the understanding of the mechanisms by which GATA3 splicing mutations can cause HDR syndrome.

Genome-wide identification, evolution and expression pattern analysis of the GATA gene family in Sorghum bicolor.

The GATA family of transcription factors is zinc finger DNA binding proteins involved in a variety of biological processes, including plant growth and development and response to biotic/abiotic stresses, and thus play an essential role in plant response to environmental changes. However, the GATA gene family of Sorghum (SbGATA) has not been systematically analyzed and reported yet. Herein, we used a variety of bioinformatics methods and quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) to explore the evolution and function of the 33 SbGATA genes identified. These SbGATA genes, distributed on 10 chromosomes, are classified into four subfamilies (I-IV) containing one pair of tandem duplications and nine pairs of segment duplications, which are more closely related to the monocot Brachypodium distachyon and Oryza sativa GATA genes. The physicochemical properties of the SbGATAs are significantly different among the subfamilies, while the protein structure and conserved protein motifs are highly conserved in the subfamilies. In addition, the transcription of SbGATAs is tissue-specific during Sorghum growth and development, which allows for functional diversity in response to stress and hormones. Collectively, our study lays a theoretical foundation for an in-depth analysis of the functions, mechanisms and evolutionary relationships of SbGATA during plant growth and development.

Genetic insights into the Tetralogy of Fallot

Tetralogy of Fallot (TOF), (OMIM #187500) one of the first known congenital heart disease (CHDs) with a rising frequency of adult patients, is a suitable paradigm for our analysis given the expanding abundance of genetic data available and these clinical consequences. Given the complexity of cardiac development, it has been associated with untreated maternal diabetes, maternal intake of retinoic acid, phenylketonuria, chromosomal anomalies (trisomies 21, 18, 13), microdeletions of chromosome 22q11.2, and it is not unexpected that a variety of transcription factors and signaling molecules involved in 9 cardiogenesis have been linked to TOF, with the literature consistently reporting the existence of new, previously unrecognized genes. The well studied genes GATA4, NKX2.5, JAG1, FOXC2, TBX5, and TBX1, which have previously been linked to TOF, are the focus of this review.

Interactive effects of light quality and nitrate supply on growth and metabolic processes in two lettuce cultivars (Lactuca sativa L.)

Light quality and nutrient parameters alter plant growth and metabolic pathways in leafy greens, therefore being primarily optimized in indoor precision agriculture. While their individual effects were studied extensively with large-scale phenomics and transcriptomics, the combinatorial effects remain poorly understood. Here, we explored the hidden interactions among light- and nitrate-responsive transcriptomes and their associations with six growth and biochemical traits, using the full factorial experimental design with three lights (red (R), blue (B) and a 1:1 mixture of red and blue (RB)), two nitrate concentrations and two cultivars (Green oak-leaf and Red oak-leaf). Multifactor analysis of 36 RNA-sequencing data revealed their largely independent but additive effects that are associated with MYB, bHLH and GATA family transcription factors. Red light and sufficient nitrate supply additively increased the primary metabolite pathway and bHLH and GATA genes, leading to a high growth rate. Blue and RB lights, together with nitrate deficiency, induced flavonoid pathway and MYB genes. Variety-specific deviations were detected in late anthocyanin biosynthetic genes and specific MYB transcription factors that would explain anthocyanin accumulations in the Red oak-leaf cultivar. Taken together, our large-scale transcriptomic and physiologic results suggest that the two environmental parameters (i.e., light quality and nitrate level) (1) utilize independent transcriptional regulatory mechanisms to achieve similar phenotypic and metabolic changes, and (2) induce the variety-specific anthocyanin accumulations via a small number of genes that show different gene expression profiles between the two cultivars.

Identification, Characterization, and Expression Profiling of Maize GATA Gene Family in Response to Abiotic and Biotic Stresses

GATA transcription factor is crucial for plant growth and development, physiological metabolism, and environmental response, which has been reported in many plants. Although the identification of maize GATA genes has been reported previously, the number of maize GATA genes was incomplete, and the expression patterns of maize GATA genes were not analyzed. Therefore, in this study, the GATA gene family of maize (Zea mays L.) was systematically analyzed. Forty-one GATA family genes were identified in the maize and were divided into four groups. The gene structure of each subgroup was basically consistent with that of the motif. The maize GATA genes were distributed on 10 chromosomes, including 3 and 17 pairs of tandem and segmental duplication genes, respectively. Fourteen types of cis-acting elements were identified in the promoter sequences of maize GATA family genes, involving four categories: light response, stress, hormone, and growth and development. The tissue-specific expression analysis of maize GATA family genes revealed that 4 GATA genes were highly expressed in almost all the maize tissues, and 11 GATA genes were not expressed in almost all tissues. The other maize GATA family genes showed a tissue-specific expression pattern. The results of RNA-seq reanalysis of publicly available transcriptome sequencing big data revealed that the gene ZmGATA37 was significantly down-regulated in response to abiotic stresses including high temperature, low temperature, drought, waterlogging, and salt, and significantly up-regulated in response to biotic stresses including smut disease, Maize Iranian mosaic virus infection, beet armyworm and aphid infestations. This indicated that the ZmGATA37 gene plays an important role in maize growth and development. Our findings offer new insight into the potential role of GATA transcription factors in abiotic and biotic stresses and provide a theoretical groundwork for the molecular mechanisms underlying maize adaptation to such stress.

Ascorbic acid and salvianolic acid B enhance the valproic acid and 5-azacytidinemediated cardiac differentiation of mesenchymal stem cells.

Cardiovascular diseases remain a major cause of death globally. Cardiac cells once damaged, cannot resume the normal functioning of the heart. Bone marrow derived mesenchymal stem cells (BM-MSCs) have shown the potential to differentiate into cardiac cells. Epigenetic modifications determine cell identity during embryo development via regulation of tissue specific gene expression. The major epigenetic mechanisms that control cell fate and biological functions are DNA methylation and histone modifications. However, epigenetic modifiers alone are not sufficient to generate mature cardiac cells. Various small molecules such as ascorbic acid (AA) and salvianolic acid B (SA) are known for their cardiomyogenic potential. Therefore, this study is aimed to examine the synergistic effects of epigenetic modifiers, valproic acid (VPA) and 5-azacytidine (5-aza) with cardiomyogenic molecules, AA and SA in the cardiac differentiation of MSCs. BM-MSCs were isolated, propagated, characterized, and then treated with an optimized dose of VPA or 5-aza for 24h. MSCs were maintained in a medium containing AA and SA for 21 days. All groups were assessed for the expression of cardiac genes and proteins through q-PCR and immunocytochemistry, respectively. Results show that epigenetic modifiers VPA or 5-aza in combination with AA and SA significantly upregulate the expression of cardiac genes MEF2C, Nkx2.5, cMHC, Tbx20, and GATA-4. In addition, VPA or 5-aza pretreatment along with AA and SA enhanced the expression of the cardiac proteins connexin-43, GATA-4, cTnI, and Nkx2.5. These findings suggest that epigenetic modifiers valproic acid and 5-azacytidine in combination with ascorbic acid and salvianolic acid B promote cardiac differentiation of MSCs. This pretreatment strategy can be exploited for designing future stem cell based therapeutic strategies for cardiovascular diseases.

New insights into triazole fungicide-caused hematopoietic abnormality in zebrafish based on GRα screening developmental toxicity

Triazole fungicides (TFs) are known to be common environmental contaminants that can be toxic to aquatic animals, but their developmental toxicity is not fully understood. To address this gap, we first used a glucocorticoid receptor α (GRα)-mediated dual luciferase reporter gene system to explore the possible development toxicity of ten TFs and found that flusilazole (FLU) exhibited stronger agonistic activity against GRα. Subsequent transcriptome sequencing showed that FLU exposure affected GRα activation and hematopoiesis associated with a variety of biological processes, including responses to corticosteroid release, embryonic hematopoiesis, erythroid differentiation, and the development of hematopoietic or lymphoid organs. Furthermore, based on in situ hybridization and staining techniques, we clarified that FLU decreased the expression of the primitive hematopoietic marker genes gata1 and pu.1. and caused the defects in the posterior blood island (PBI), thereby impacting intermediate hematopoietic processes. Also, FLU significantly reduced the expression of the crucial hematopoietic gene cmyb and disrupted the production of erythrocytes and bone marrow cells during definitive hematopoiesis. Consistently, we found that FLU induced lesions in the kidney, a hematopoietic organ, including the infiltration of inflammatory cells, tubular collapse, reduced tubular filtration area, and interstitial hydronephrosis. We also found that FLU increased aberrant red blood cells in the peripheral blood of zebrafish. These findings provide new insights into the developmental toxicity and ecotoxicological risk of TFs.

Exposure to manganese (II) chloride induces developmental toxicity, oxidative stress and inflammatory response in Marine medaka (Oryzias melastigma) embryos

Manganese (Mn) is an essential metal for organisms, but high levels can induce serious toxicity. To date, the toxic mechanism of Mn to marine fish is still poorly understood. In the present study, Oryzias melastigma embryos were exposed to different concentrations of MnCl2 (0–152.00 mg/L) to investigate its effect on early development. The results showed that exposure to MnCl2 caused developmental toxicity to embryos, including increased heart rate, delayed hatching time, decreased hatching rate and increased malformation rate. MnCl2 exposure could induce oxidative stress in O. melastigma embryos, as indicated by increased the contents of malondialdehyde (MDA) and the activities of the antioxidant enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT)). The heart might be an important target organ for MnCl2 because of cardiac malformations and disruption in the expression of cardiac development-related genes (ATPase, epo, fg8g, cox1, cox2, bmp4 and gata4). In addition, the expression levels of stress- (omTERT and p53) and inflammation-related genes (TNFα and il1β) were significantly up-regulated, suggesting that MnCl2 can trigger stress and inflammatory response in O. melastigma embryos. In conclusion, this study demonstrated that MnCl2 exposure can induce developmental toxicity, oxidative stress and inflammatory response in O. melastigma embryos, providing insights into the toxic mechanism of Mn to the early development of marine fish.

Myeloid Leukemia of Down Syndrome.

Myeloid leukemia of Down syndrome (ML-DS) is characterized by a distinct natural history and is classified by the World Health Organization (WHO) as an independent entity, occurring with unique clinical and molecular features. The presence of a long preleukemic, myelodysplastic phase, called transient abnormal myelopoiesis (TAM), precedes the initiation of ML-DS and is defined by unusual chromosomal findings. Individuals with constitutional trisomy 21 have a profound dosage imbalance in the hematopoiesis-governing genes located on chromosome 21 and thus are subject to impaired fetal as well as to neonatal erythro-megakaryopoiesis. Almost all neonates with DS develop quantitative and morphological hematological abnormalities, yet still only 5-10% of them present with one of the preleukemic or leukemic conditions of DS. The acquired mutations in the key hematopoietic transcription factor gene GATA1, found solely in cells trisomic for chromosome 21, are considered to be the essential step for the selective growth advantage of leukemic cells. While the majority of cases of TAM remain clinically 'silent' or undergo spontaneous remission, the remaining 20% to 30% of them progress into ML-DS until the age of 4 years. The hypersensitivity of ML-DS blasts to chemotherapeutic agents, including but not limited to cytarabine, and drugs' increased infectious and cardiac toxicity have necessitated the development of risk-adapted treatment protocols for children with ML-DS. Recent advances in cytogenetics and specific molecular mechanisms involved in the evolution of TAM and ML-DS are reviewed here, as well as their integration in the improvement of risk stratification and targeted management of ML-DS.

Systematic Characterization of GATA Transcription Factors in Liriodendron chinense and Functional Validation in Abiotic Stresses.

The Liriodendron chinense in the Magnoliaceae family is an endangered tree species useful for its socio-economic and ecological benefits. Abiotic stresses (cold, heat, and drought stress), among other factors, affect its growth, development, and distribution. However, GATA transcription factors (TFs) respond to various abiotic stresses and play a significant role in plant acclimatization to abiotic stresses. To determine the function of GATA TFs in L. chinense, we investigated the GATA genes in the genome of L. chinense. In this study, a total of 18 GATA genes were identified, which were randomly distributed on 12 of the total 17 chromosomes. These GATA genes clustered together in four separate groups based on their phylogenetic relationships, gene structures, and domain conservation arrangements. Detailed interspecies phylogenetic analyses of the GATA gene family demonstrated a conservation of the GATAs and a probable diversification that prompted gene diversification in plant species. In addition, the LcGATA gene family was shown to be evolutionarily closer to that of O. sativa, giving an insight into the possible LcGATA gene functions. Investigations of LcGATA gene duplication showed four gene duplicate pairs by the segmental duplication event, and these genes were a result of strong purified selection. Analysis of the cis-regulatory elements demonstrated a significant representation of the abiotic stress elements in the promoter regions of the LcGATA genes. Additional gene expressions through transcriptome and qPCR analyses revealed a significant upregulation of LcGATA17, and LcGATA18 in various stresses, including heat, cold, and drought stress in all time points analyzed. We concluded that the LcGATA genes play a pivotal role in regulating abiotic stress in L. chinense. In summary, our results provide new insights into understanding of the LcGATA gene family and their regulatory functions during abiotic stresses.

Genomic characterization of the GATA3 mutational landscape in breast cancer.

1039 Background: GATA3 expression is broadly used as a biomarker for diagnosing cancer of breast origin and its expression is strongly associated with estrogen receptor (ER)-positive luminal phenotype. Although GATA3 mutations are observed in 12-18% of breast cancers (BC), they are poorly characterized. Recently, the pharmacological inhibition of MDM2 has been shown to significantly impair tumor growth in GATA3-deficient models in vitro, in vivo, and patient-derived xenograft harboring GATA3 somatic mutations. Therefore, given the potential targetability of GATA3-mutated BC cells, it is important to better understand and characterize the mutational landscape of GATA3 to help guide future prospective clinical studies. Methods: We accessed a cohort of BC samples profiled genomically and transcriptomically from two open-source datasets: TCGA (n=961) and METABRIC (n=1866). We used the chi-squared test to analyze the frequency of GATA3 mutations across PAM50 subtypes (Basal, HER2, Luminal A, Luminal B, Normal-like) and histological BC subtypes (invasive ductal carcinoma [IDC] and invasive lobular carcinoma [ILC]). Mutations affecting other genes in the GATA3-mutated samples and the GATA3 wild-type (WT) samples were compared using the chi-squared test. We analyzed the mutational landscape of the GATA3 gene and correlated different sites of mutations with GATA3 expression, MDM2 amplification, other co-occurring mutations, and clinical behavior. GATA3 expression levels were compared using the Mann-Whitney test. Results: Of the analyzed 2,827 BC samples, GATA3 mutations were unevenly distributed in the five PAM50 subtypes, as Luminal A and B subtypes had the highest frequency (16% and 17%, respectively) and the basal subtype had no observed GATA3 mutations (p <0.0001). Further analysis showed that GATA3 mutations were more common in luminal IDC than ILC (17% vs 8.7%; p <0.0001). While mutations in CBFB, AKT1, TBX3 and ARID1A were more frequently co-occurring with GATA3 mutations, mutations in TP53, CDH1 and PIK3CA were mutually exclusive with GATA3 mutations (adjusted p <0.0001 for all genes). Analysis of the GATA3 mutational landscape identified two types of mutations in GATA3: a group of truncating mutations occurring at the end of the GATA3 gene (after the 307 position including the splice site at 308) and a group occurring before the 307 position. The first group was associated with increased GATA3 expression, MDM2 amplification, and was mutually exclusive with TP53 mutations. The second group had little to no effect on GATA3 expression and behaved similarly to GATA3 WT. Conclusions: A subtype of GATA3 mutations are mutually exclusive with TP53 mutations and are associated with increased MDM2 amplification, making them an ideal target for clinical trials involving MDM2 inhibitors.

Identification of a novel splice site variant in the GATA3 gene in a Chinese patient with hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome, and a review of genotype-phenotype spectrum

Identification of a novel splice site variant in the GATA3 gene in a Chinese patient with hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome, and a review of genotype-phenotype spectrum

Novel urine-based DNA methylation biomarkers for urothelial bladder carcinoma detection in patients with hematuria

ABSTRACT Background Urothelial bladder carcinoma (UBC) is usually detected during work-up for hematuria. Cystoscopy and/or contrast-enhanced imaging are the gold standard tools for UBC diagnosis, despite limited by being invasive, expensive and low yield in small flat tumors. Objectives To assess the diagnostic performance of urine-based DNA methylation of six genes (GATA4, P16, P14, APC, CDH1 and CD99) for UBC detection in patients with hematuria. Patients and methods Voided urine was collected from consecutive patients presented with hematuria for urine cytology and DNA methylation assay of the assigned genes using methylation-specific Polymerase Chain Reaction (PCR). Further assessment by office cystoscopy and imaging with subsequent inpatient cystoscopic biopsy for positive findings was done. The diagnostic characteristics of DNA methylation and urine cytology were assessed based on its capability to predict UBC. Results We included 246 patients in the study with identified macroscopic hematuria in 204 (82.9%) patients. Positive cytology was found in 78 (31.7%) patients. DNA methylation of GATA4, P16, P14, APC, CDH1 and CD99 genes was identified in 127 (51.6%), 52 (21.1%), 117 (47.6%), 106 (43.1%), 90 (36.6%) and 71 (28.9%) patients, respectively. The sensitivity of the assigned genes for UBC detection ranges from 35% (95%CI: 31–39) to 83% (95%CI: 79–87). Optimal specificity (SP) (100%) was noted for P16, APC and CDH1 genes. While for the other genes (GATA4, P14 and CD99), the SP was 95% (95%CI: 92–98), 96% (95%CI: 92–99) and 97% (95%CI: 93–99), respectively. On multivariate logistic regression analysis, all genes exclusively demonstrated independent prediction of UBC. On receiver operator characteristic (ROC) analysis, all tested genes methylation showed superior area under the curve (AUC) when compared to urine cytology. Conclusions We have developed a novel urine-based DNA methylation assay for detection of UBC in patients with hematuria with superior diagnostic performance and independent predictive capacity over urine cytology.

The Alleviating Impacts of Quercetin on Inflammation and Oxidant-antioxidant Imbalance in Rats with Allergic Asthma.

Asthma is an inflammatory disease of the airways. We assessed the anti-inflammatory and antioxidative impacts of quercetin, a plant derivative, on inflammatory and oxidative indices in lung tissue and serum of rats with asthma.Asth ma was induced by ovalbumin. Rats were divided into 4 groups: control, asthma+vehicle (Receieved normal saline), asthma+dexamethasone, and asthma+quercetin. After asthma induction, quercetin (50 mg/kg) and dexamethasone (2.5 mg/kg) were injected intraperitoneally once daily for 1 week. On day 50, lung histopathology indices; inflammatory factors; tissue gene expression, including GATA Binding Protein 3 (Gata-3), Tbx21 (T-bet), Transforming growth factor-β (TGF-β), Il10 (IL-10), Il1b (IL-1β), Il6 (IL-6), Acta2 (α-SMA), and Tnf (TNF-α); and oxidative stress indices (malondialdehyde [MDA], catalase [CAT], glutathioneperoxidase [GPX], superoxide dismutase [SOD], and total antioxidant capacity [TAC]) in tissue and serum, were evaluated. The results showed that quercetin reduced Gata3, Tnf, Tgfb1, Il1b, and Acta2 gene expression and increased Tbx21 gene expression following asthma. Quercetin also improved oxidative stress by decreasing MDA levels and increasing TAC, CAT, SOD, and GPX levels in serum and lung tissue. Furthermore, quercetin decreased IL6 and TNFα levels and increased IL10 levels in lung tissue after asthma was treated with quercetin. Quercetin ameliorates oxidative stress and inflammation caused by asthma, especially at the tissue level. Therefore, quercetin can be considered a potent antiasthmatic agent.

Recent progress in analyses of GATA1 in hematopoietic disorders: a mini-review

GATA1 is an essential master regulator of erythropoiesis and megakaryopoiesis. Accumulating lines of evidence have shown that dynamic changes in GATA1 gene expression levels during erythropoiesis are crucial for proper erythroid differentiation. Since GATA1 is an X-chromosome gene, GATA1 knockout leads to embryonic lethal dyserythropoiesis in male mice, while heterozygous female mice can survive. In the past decade, it has become clear that germline GATA1 gene mutations leading to structural changes in the GATA1 protein are involved in congenital dyserythropoiesis in males. In contrast, decreased GATA1 expression levels, which cause embryonic lethal dyserythropoiesis in male mice, increase the risk of erythroleukemia development in female mice, while female GATA1-knockout mice do not show substantial phenotypic alterations in erythroid or megakaryocyte lineages. In this review, we summarize the recent progress in elucidating the roles of GATA1 in normal and pathogenetic erythropoiesis and discuss the possible mechanisms of pathogenesis of dyserythropoiesis and erythroleukemia.

ACase of GATA2 Deficiency with Pancytopenia and Recurrent Infections: To Transplant or Not to Transplant?

BackgroundGATA2 deficiency is an autosomal dominant disorder arising from various mutations in the GATA2 gene, which encodes a zinc finger transcription factor that is required for hematopoiesis and angiogenesis. Patients with GATA2 deficiency can present with a range of hematologic and immunologic manifestations. We present a case of GATA2 deficiency with recurrent transient pancytopenia who is also heterozygous for C9 deficiency. Case presentationOur 30-year-old female patient presented with febrile neutropenia (ANC 850) and rash. Interestingly, she reported a history of recurrent viral upper respiratory infections since childhood and had mononucleosis at the age of 5. In her mid-twenties, she started having recurrent sinusitis and pneumonias, requiring hospitalization and antibiotic therapy 1–2 times per year. She also had persistent pancytopenia for at least 5 years that transiently improved with transfusions. CBC ranged from Hgb 5.5–10.1 g/dL (mean 8), platelets 30–102 K/uL (mean 67), WBC 1.4–5.6 K/uL (mean 2.87), neutrophils 0.75–4.35 K/uL (mean 2), monocytes 0–0.07 K/uL (mean 0.02), lymphocytes 0.38–1.18 K/uL (mean 0.8). Bone marrow biopsy showed severely hypocellular bone marrow with trilineage hypoplasia without dysplasia or neoplasia. We initiated an immune workup that showed normal IgG, IgA and IgM levels; low B cell numbers; absent NK cells (CD56+); and preserved T cell numbers on flow cytometry. Strep pneumoniae IgG levels were nonprotective with less than 0.1 ug/mL for all serotypes. Genetic panel testing for primary immunodeficiency revealed a heterozygous pathogenic mutation in GATA2 (c.1081C>T (p.Arg361Cys). Upon multidisciplinary discussion with Hematology, decision was made to begin immunoglobulin replacement. Bone marrow transplant workup was initiated, but the team is hesitant to proceed with transplant at this time. DiscussionHematopoietic stem cell transplantation is the only treatment option with curative potential for this patient, but there are significant associated morbidity and mortality risks. Identifying the appropriate timing for transplantation remains a central focus of this patient's care. Close monitoring of the patient's overall clinical evolution including peripheral cell counts, bone marrow findings, and infection frequency and severity could help approximate the optimal transplantation timing after careful consideration of risks and benefits.

Barakat syndrome: clinical polymorphism of the disease

Barakat Syndrome (MIM#146255) is a rare autosomal dominant disease caused by GATA3 gene mutation and manifested by hypoparathyroidism (H), sensorineural deafness (D), and renal disease (R). HDR syndrome characterized by high clinical variability and prognosis. The exact prevalence of this disease is unknown, 180 cases are reported in the literature. Two clinical cases are presented. De novo heterozygous mutation in the GATA3 gene was detected in both patients. Our observations demonstrate variability of clinical phenotypes and poor prognosis in patients with Barakat Syndrome. The syndrome should be suspected in cases of early high-grade deafness and kidney disease presentation for the purpose of early diagnosis and appropriate therapy including the prevention of CKD progression.

Successful allogeneic hematopoietic stem cell transplantation for myelodysplastic neoplasms complicated with secondary pulmonary alveolar proteinosis and Behçet's disease harboring GATA2 mutation.

Myelodysplastic neoplasms (MDS) are defined by cytopenia and morphologic dysplasia originating from clonal hematopoiesis. They are also frequently complicated with diseases caused by immune dysfunction, such as Behçet's disease (BD) and secondary pulmonary alveolar proteinosis (sPAP). MDS with both BD and sPAP is extremely rare, and their prognosis is poor. In addition, haploinsufficiency of the hematopoietic transcription factor gene GATA2 is recognized as a cause of familial MDS and is frequently complicated by sPAP. Herein, we report a case of MDS combined with both BD and sPAP in association with GATA2 deficiency in a Japanese woman. Because she developed progressive leukopenia and macrocytic anemia during BD treatment at the age of 61, she underwent a bone-marrow examination and was diagnosed with MDS. She subsequently developed sPAP. At the age of 63, she underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT). Since allo-HSCT, she has maintained complete remission of MDS as well as the symptoms of BD and sPAP. Furthermore, we performed whole exome sequencing and identified the GATA2 Ala164Thr germline mutation. These findings suggest that patients with MDS, BD and sPAP should be considered for early allo-HSCT.

Integrated analysis of transcriptome-wide m6A methylation in a Cd-induced kidney injury rat model

BackgroundAccumulating evidence has demonstrated that N6-methyladenosine (m6A) plays important roles in a variety of diseases. However, the specific functions of m6A in CdCl2-induced kidney injury remain unclear. ObjectiveHere, we investigate a transcriptome-wide map of m6A modifications and explore the effects of m6A on Cd-induced kidney injury. Materials and methodsThe rat kidney injury model was constructed by subcutaneous injection of CdCl2 (0.5, 1.0, and 2.0 mg/kg). The m6A levels were measured by colorimetry. The level of expression of m6A-related enzymes were detected by reverse transcription quantitative real-time PCR analysis. Transcriptome-wide m6A methylome in CdCl2 (2.0 mg/kg) and the control group were profiled by methylated RNA immunoprecipitation sequencing (MeRIP-seq). Subsequently, the sequencing data were analyzed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG), while gene set enrichment analysis (GSEA) confirmed the functional enrichment pathways of sequencing genes. In addition, a protein-protein interaction (PPI) network was applied to select hub genes. ResultsThe levels of m6A and m6A regulators (METTL3, METTL14, WTAP, YTHDF2) were significantly increased in CdCl2 groups. We identified a total of 2615 differentially expressed m6A peaks, 868 differentially expressed genes and 200 genes with significant changes in both m6A modification and gene expression levels. GO, KEGG, and GSEA analyses indicated that these genes were mainly enriched in inflammation and metabolism-related pathways such as in IL-17 signaling and fatty acid metabolism. According the result of the conjoint analysis, we identified the top ten hub genes (Fos, Hsp90aa1, Gata3, Fcer1g, Cftr, Cspg4, Atf3, Cdkn1a, Ptgs2, and Npy) which may be regulated by m6A and involve in CdCl2-induced kidney damage. ConclusionThis study established a m6A transcriptional map in a CdCl2-induced kidney injury model and suggested that m6A may affect CdCl2 induced kidney injury via regulated the inflammation and metabolism related gene.

Abstract 5923: The genetic landscape of metastatic breast cancer reflected in circulating tumor DNA

Abstract Tissue biopsies of breast tumors only provide a snapshot of the evolution of the disease and may miss potential therapeutic targets, especially in the metastatic setting. Plasma-derived cell-free tumor DNA constitutes a potential surrogate for tumor DNA obtained from tissue biopsies to potentially capture inter-and intra-tumoral heterogeneity present in metastatic breast cancer. We aimed to investigate to what extent variants identified in the primary tumor and metastases are reflected in plasma. We performed whole-exome sequencing on tumor DNA from primary and metastatic lesions and plasma-derived circulating tumor DNA of eight patients with metastatic breast cancer. Our results show that metastases most often follow parallel evolution, given the low levels of stem mutations detected between primary tumors and matched metastases and mutational events private to the primary tumor. Furthermore, we demonstrated that ctDNA mainly reflected old stem mutations shared between primary tumor and metastasis, and less often, genetic alterations found solely in primary tumor or metastasis. Lastly, clinically relevant variants, e.g., resistance mutations detected in primary tumor and metastasis, were to some extent reflected in ctDNA, including affected genes MET, MTOR, CDK6, PIK3CA, and GATA3. However, mutations in MED1, NCOR1, and NCOR2 were only found in tumor tissues. These findings support the importance of a metastatic deposit biopsy to guide treatment decisions. Together, our results have implications for future ctDNA studies. For monitoring tumor burden using ctDNA, our results suggest that truncal mutations are the best candidates, as they are highest in circulating levels. For confident tracking of subclonal mutations in ctDNA, liquid biopsies are not yet to substitute metastatic tissue biopsies and instead demonstrate support to the current gold standard. Citation Format: Stephanie Kavan. The genetic landscape of metastatic breast cancer reflected in circulating tumor DNA [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5923.