Research progress in role and mechanism of YAP/TAZ in the development of renal cell carcinoma / 中国肿瘤生物治疗杂志

@#[摘 要] 肾癌是泌尿系统最常见的肿瘤之一，早期临床表现不典型，经手术治疗后仍有部分发生复发和转移，病死率高。因此，肾癌的早期诊断和晚期治疗成为亟待解决的两大问题。以YAP/TAZ为核心的Hippo通路在肾癌的发生发展中发挥着重要作用，YAP/TAZ通过直接调控肾癌细胞的转录活性、内源性竞争RNA（ceRNA）机制、促进血管形成、增强肾细胞铁死亡敏感性以及作为肾癌细胞中其他信号通路的转导枢纽等多种机制，促进肾癌细胞的侵袭、转移和耐药；除此之外，YAP/TAZ在多种罕见的肾癌组织学亚型中起到指导分型和预测预后的作用。对于YAP/TAZ在肾癌中的作用及其机制的认识可为临床肾癌的早期诊断和晚期治疗提供理论参考依据。

NDFIP1 limits cellular TAZ accumulation via exosomal sorting to inhibit NSCLC proliferation

NDFIP1 has been previously reported as a tumor suppressor in multiple solid tumors, but the function of NDFIP1 in NSCLC and the underlying mechanism are still unknown. Besides, the WW domain containing proteins can be recognized by NDFIP1, resulted in the loading of the target proteins into exosomes. However, whether WW domain-containing transcription regulator 1 (WWTR1, also known as TAZ) can be packaged into exosomes by NDFIP1 and if so, whether the release of this oncogenic protein via exosomes has an effect on tumor development has not been investigated to any extent. Here, we first found that NDFIP1 was low expressed in NSCLC samples and cell lines, which is associated with shorter OS. Then, we confirmed the interaction between TAZ and NDFIP1, and the existence of TAZ in exosomes, which requires NDFIP1. Critically, knockout of NDFIP1 led to TAZ accumulation with no change in its mRNA level and degradation rate. And the cellular TAZ level could be altered by exosome secretion. Furthermore, NDFIP1 inhibited proliferation in vitro and in vivo, and silencing TAZ eliminated the increase of proliferation caused by NDFIP1 knockout. Moreover, TAZ was negatively correlated with NDFIP1 in subcutaneous xenograft model and clinical samples, and the serum exosomal TAZ level was lower in NSCLC patients. In summary, our data uncover a new tumor suppressor, NDFIP1 in NSCLC, and a new exosome-related regulatory mechanism of TAZ.

A YAP/TAZ-CD54 axis is required for CXCR2-CD44- tumor-specific neutrophils to suppress gastric cancer

As an important part of tumor microenvironment, neutrophils are poorly understood due to their spatiotemporal heterogeneity in tumorigenesis. Here we defined, at single-cell resolution, CD44-CXCR2- neutrophils as tumor-specific neutrophils (tsNeus) in both mouse and human gastric cancer (GC). We uncovered a Hippo regulon in neutrophils with unique YAP signature genes (e.g., ICAM1, CD14, EGR1) distinct from those identified in epithelial and/or cancer cells. Importantly, knockout of YAP/TAZ in neutrophils impaired their differentiation into CD54+ tsNeus and reduced their antitumor activity, leading to accelerated GC progression. Moreover, the relative amounts of CD54+ tsNeus were found to be negatively associated with GC progression and positively associated with patient survival. Interestingly, GC patients receiving neoadjuvant chemotherapy had increased numbers of CD54+ tsNeus. Furthermore, pharmacologically enhancing YAP activity selectively activated neutrophils to suppress refractory GC, with no significant inflammation-related side effects. Thus, our work characterized tumor-specific neutrophils in GC and revealed an essential role of YAP/TAZ-CD54 axis in tsNeus, opening a new possibility to develop neutrophil-based antitumor therapeutics.

The combination of YAP/TAZ predicts the clinical prognosis in patients with cholangiocarcinoma after radical resection / 中华内科杂志

Objective:Cholangiocarcinoma (CCA) is a malignant tumor derived from bile duct epithelial cells with extremely poor prognosis. The Hippo-Yes-associated protein (YAP)/transcription activator with PDZ binding motif (TAZ) signaling plays a critical role in cancer stem cell biology. Previous studies have shown that the positive expression of YAP/TAZ in CCA predicts larger tumor size and unfavorable clinical outcomes. We aim to evaluate the prognostic value of YAP/TAZ detection in CCA patients.Methods:CCA patients who underwent radical resection were retrospectively analyzed at our institution from January 2011 to June 2016. Postoperative pathological specimens were scored by YAP/TAZ immunohistochemical staining. The prognostic value of YAP/TAZ was analyzed by multivariate Cox-proportional hazards model.Results:A total of 91 CCA patients were enrolled. During a median follow-up time of 11.0 months, 69.2% patients relapsed and 45.1% died. The median OS and DFS were 10.7 months and 8.8 months respectively. The YAP/TAZ dual positive patients owned a worse TNM stage ( P=0.015), poorer tissue differentiation ( P=0.007), and a higher CA199 than those in negative patients. Multivariate Cox analysis identified that YAP/TAZ dual positivity as a significant factor predicted poorer OS ( P=0.010) and DFS ( P=0.028) in CCA patients after radical resection. In subgroup analysis, YAP/TAZ combination also significantly predicted OS ( P=0.044) and DFS ( P=0.043) in CCA patients with positive lymphatic metastasis and/or surgical margin who required adjuvant therapy. Conclusions:YAP/TAZ positivity is an independent predictive factor for survival in CCA patients after radical resectiony.

Targeting-YAP/TAZ therapies for head and neck cancer, directly or indirectly? / 华西口腔医学杂志

YAP/TAZ are wild over-activated in head and neck squamous cell carcinoma (HNSCC) with high potential as a direct therapy target for HNSCC treatments. However, the efforts on the directly targeting-YAP/TAZ therapies over the past decade, have very limited impacts, mainly caused by: 1. There is still none effective and specific YAP/TAZ inhibitor with clinical potential; 2. YAP/TAZ might not be directly targeted, because of their multiple important biological functions, such as: regulation of cell proliferation and survival, stem cell maintain, regulation of organ development, organ size control, and tissue regeneration. Interestingly, the over-activation of YAP/TAZ in HNSCC mainly be regulated by upstream abnormal molecular or biological events, instead of genes alteration of YAP/TAZ. Therefore, exploring the alternative molecular events regulating YAP/TAZ activation and molecular mechanism in HNSCC might help to uncover novel indirect targets of YAP/TAZ therapies for HNSCC prevention and treatment.

Correlation between Notch signaling pathway and YAP1/TAZ in breast cancer / 西安交通大学学报(医学版)

Objective: To investigate the relationship of Notch1 expression with YAP1/TAZ expression in breast cancer and its possible mechanism. Methods: MDA-MB-231 and MCF-7 breast cancer cells were selected, and lentiviruses were used to construct stable infected cell lines with different expression levels of Notch1 and YAP1. Western blotting, RT-qPCR and co-immunoprecipitation were used to study the relationship between Notch1 and YAP1/TAZ expressions. Results: On the gene expression profiling interactive analysis (GEPIA) website, we analyzed the breast cancer data in the Cancer Gene Atlas Database (TCGA) and found that the expressions of Notch1 and YAP1/TAZ were positively correlated. Compared with those in the control group, the protein and mRNA levels of YAP1/TAZ in the shNotch1 group were reduced (P0.05). Co-immunoprecipitation showed that TAZ protein interacted with Notch1 and β-TrCP protein. Compared with those in the control group, the mRNA and protein levels of Notch1 and JAG1 in shYAP1 group were reduced (P<0.05), while those in YAP1 group were increased (P<0.05). TEAD family was predicted to be a JAG1 transcription factor on the JASPAR 2018 website. TEAD1/3/4 siRNA could effectively inhibit TEAD1/3/4 expression, and JAG1 expression decreased too (P<0.05). Conclusion: There is a positive feedback loop between Notch1 and YAP1/TAZ in breast cancer. YAP1/TAZ-TEAD can activate the Notch1 signaling pathway by regulating JAG1 expression. Notch1 protein can affect the degradation of YAP1/TAZ protein.

Yes-associated protein (YAP) and transcriptional coactivator with a PDZ-binding motif (TAZ): a nexus between hypoxia and cancer

Hypoxia is a common feature of solid tumors. As transcription factors, hypoxia-inducible factors (HIFs) are the master regulators of the hypoxic microenvironment; their target genes function in tumorigenesis and tumor development. Intriguingly, both yes-associated protein (YAP) and its paralog transcriptional coactivator with a PDZ-binding motif (TAZ) play fundamental roles in the malignant progression of hypoxic tumors. As downstream effectors of the mammalian Hippo pathway, YAP and/or TAZ (YAP/TAZ) are phosphorylated and sequestered in the cytoplasm by the large tumor suppressor kinase 1/2 (LATS1/2)-MOB kinase activator 1 (MOB1) complex, which restricts the transcriptional activity of YAP/TAZ. However, dephosphorylated YAP/TAZ have the ability to translocate to the nucleus where they induce transcription of target genes, most of which are closely related to cancer. Herein we review the tumor-related signaling crosstalk between YAP/TAZ and hypoxia, describe current agents and therapeutic strategies targeting the hypoxia-YAP/TAZ axis, and highlight questions that might have a potential impact in the future.

Tumor suppressive function of Matrin 3 in the basal-like breast cancer

BACKGROUND: Basal-like breast cancer (BLBC) or triple-negative breast cancer (TNBC) is an aggressive and highly metastatic subtype of human breast cancer. The present study aimed to elucidate the potential tumor-suppressive function of MATR3, an abundant nuclear protein, in BLBC/TNBC, whose cancer-relevance has not been characterized. METHODS: We analyzed in vitro tumorigenecity by cell proliferation and soft agar colony formation assays, apoptotic cell death by flow cytometry and Poly (ADP-ribose) polymerase (PARP) cleavage, epithelial-mesenchymal transition (EMT) by checking specific EMT markers with real-time quantitative PCR and in vitro migration and invasion by Boyden Chamber assays. To elucidate the underlying mechanism by which MATR3 functions as a tumor suppressor, we performed Tandem affinity purification followed by mass spectrometry (TAP-MS) and pathway analysis. We also scrutinized MATR3 expression levels in the different subtypes of human breast cancer and the correlation between MATR3 expression and patient survival by bioinformatic analyses of publicly available transcriptome datasets. RESULTS: MATR3 suppressed in vitro tumorigenecity, promoted apoptotic cell death and inhibited EMT, migration, and invasion in BLBC/TNBC cells. Various proteins regulating apoptosis were identified as MATR3-binding proteins, and YAP/TAZ pathway was suppressed by MATR3. MATR3 expression was inversely correlated with the aggressive and metastatic nature of breast cancer. Moreover, high expression levels of MATR3 were associated with a good prognosis of breast cancer patients. CONCLUSIONS: Our data demonstrate that MATR3 functions as a putative tumor suppressor in BLBC/TNBC cells. Also, MATR3 potentially plays a role as a biomarker in predicting chemotherapy-sensitivity and patient survival in breast cancer patients.

TAZ promoting angiogenesis and its mechanism in gastric cancer / 中国肿瘤临床

Objective: To determine the expression of TAZ and its role in angiogenesis in gastric carcinoma. Methods: Immunohistochemical staining was performed to investigate the expression of TAZ and to determine whether a direct relationship exists between TAZ and β-catenin. Transfection with TAZ overexpression plasmid in MKN28 cells was conducted to induce exogenous expression of TAZ and a TAZ knockdown plasmid was transfected into MGC803 cells to reduce TAZ levels. The effects on endothelial cell formation, proliferation, and migration were determined by Matrigel three-dimensional culture, MTT proliferation assay and Transwell migration assay. In addition, the expression of TAZ and β-catenin in transfected gastric cancer cells was detected by Western blot. Results: Immunohistochemistry showed that TAZ protein was expressed in 64 of 150 gastric cancer sample tissues (43%), TAZ was localized in the nucleus, and its expression was associated with tumor grade, TNM stage, metastasis, and microvessel density (MVD) (P<0.05). In addition, the expression frequency of β-catenin in the TAZ positive group was 67.2%, which was significantly higher than that in the TAZ negative group, and the expression of TAZ was positively correlated with β-catenin. After transfection, TAZ overexpression increased the expression of β-catenin and enhanced HUVECs tube formation, proliferation, and migration. In the MGC803 cells transfected with the knockdown plasmid, β-catenin levels were decreased and HUVECs motility was inhibited. Conclusions: TAZ may promote angiogenesis in gastric cancer by promoting β-catenin expression.

Pepstatin Pr show anti-fibrosis effect related to YAP-TGFβ-Smad pathway / 药学学报

Liver fibrosis is a tissue repair compensatory response to liver injury caused by various chronic factors, ultimately leading to liver cirrhosis, liver failure and even hepatocellular carcinoma. Abnormal activation of hepatic stellate cells is the cellular basis of liver fibrosis development. Pepstatin Pr, the derivative of pepstatin A, was isolated from Streptomyces sp. CPCC 202950. Our purpose was to investigate the anti-fibrotic activity of pepstatin Pr and explore its molecular mechanism. Hepatic stellate cell LX-2 was stimulated by TGFβ1 and sub- sequently treated with pepstatin Pr. Its cytotoxicity was detected by sulforhodamine B (SRB) assay. The expression of COL1A1, α-SMA and cathepsin D, signaling proteins TGFβ, Smad and YAP/TAZ were detected by Western blot or real-time PCR. The results showed that pepstatin Pr was not cytotoxic to LX-2 cells. And pepstatin Pr significantly reduced the mRNA and protein expression of COL1A1 and α-SMA, which are important liver fibrosis markers. Pepstatin Pr also repressed the protein expression level of cathepsin D, TGFβ1, YAP/TAZ, the phospholation level of Smad2, and YAP nuclear translocation. In conclusion, pepstatin Pr exhibits anti-fibrotic effects in TGFβ1-stimulaed LX-2 cells by mediating YAP-TGFβ-Smad pathway.

Expression of YAP, LPA and TAZ in head and neck squamous cell carcinoma and its correlation with clinicopathological data / 实用医学杂志

Objective To investigate the expression of YAP,LPA and TAZ in head and neck squamous cell carcinoma and its clinical significance.Methods Selective collection of Ya'an People's Hospital pathology department from January 2010 to December 2016,the surgical resection of the pathological tissue paraffin specimens amounted to 136 cases,including the normal tissue of head and neck squamous cell carcinoma 37 cases,head and neck benign tumor-like tissue 35 cases,head and neck squamous cell carcinoma 64 cases,all specimens without chemotherapy history,The relationship between the expression of LPA,YAP and TAZ in various tissues by immunohistochemistry and real-time quantitative PCR assay and its clinical correlative parameters.Results The expression of LPA mRNA in normal tissues,benign neoplasia tissues and squamous carcinoma tissues of the head and neck squamous cell carcinoma was similar (P ＞ 0.05).The expression of YAP mRNA in squamous cell carcinoma tissues was higher than that in the adjacent tissues and tumor tissues (P ＜ 0.05).TAZ mRNA was low expression in the three tissues of the head and neck,but the expression of squamous cell carcinoma was significantly higher than that of benign tumor tissues and adjacent normal tissues (P ＜ 0.05).The expression of YAP and TAZ protein was significantly higher than that in the tissues of normal tissues and benign tumor tissues (P ＜ 0.05).The expression of YAP in squamous cell carcinoma was significantly higher than the control group (P ＜ 0.05).The expression of TAZ was low differentiation in tumor,tumor ＞3.0 cm,lymph node positive and Ⅲ-Ⅳ period were higher than the control group (P ＜ 0.05).Condusions The expression of YAP and TAZ in head and neck squamous cell carcinoma was closely related to tumor staging,differentiation degree and lymph node metastasis,while the expression of LPA showed no significant difference and correlation.

YAP/TAZ regulates multiple signal pathways in the genesis and development of hepatic fibrosis / 中南大学学报(医学版)

Hepatic fibrosis is a repair response to liver injury,and hepatic stellate cell activation is the center of hepatic fibrosis,which involves Hippo,Notch,Wnt/β-catenin,and TGF-β/Smad signaling pathways.YAP/TAZ is an important nucleus factor for Hippo tumor suppressor pathway and its activity is the key to the growth of whole organs,cell proliferation,and specific amplification of progenitor cells in the process of tissue renewal and regeneration.As the hub of signaling pathways,such as Hippo,Notch,Wnt/β-catenin,TGF-β/Smad signaling,YAP/TAZ regulates the genesis and development of liver fibrosis.

Research advances in the role of the Hippo-YAP/TAZ signaling pathway in primary liver cancer / 中华肝脏病杂志

The Hippo-YAP/TAZ signaling pathway is an evolutionarily conserved pathway, which has been confirmed to play an important role in organ volume control, stem cell function, tissue regeneration, and tumorigenesis. Recent research findings show that the Hippo-YAP/TAZ signaling pathway is closely associated with the development and progression of primary liver cancer, and inhibition of the activity of this pathway may be a new method for the treatment of liver cancer. This article reviews the research advances in the role of the Hippo-YAP/TAZ signaling pathway in primary liver cancer.

Relationship of TAZ expression between peripheral blood and tumor tissue in patients with colorectal cancer and its clinical significance / 重庆医学

Objective To investigate the expression of TAZ in peripheral blood and tumor tissue of the patients with colorectal cancer and to analyze their relationship and clinical significance.Methods Eighty-five cases of pathologically verified colorectal cancer treated by radical operation were selected as the operation group,and 35 cases with postoperative relapse served as the recurrent group.Meanwhile 50 age and sex-matched healthy persons were selected as the control group.The immunohistochemical(SP)technique was adopted to detect the expression of TAZ in tumor tissues and expression of TAZ in peripheral blood was tested with enzyme linked immunosorbent assay(ELISA).Results The level of preoperative TAZ in peripheral blood of the operation group was significantly higher than that of the control group(P＜0.05).The level of peripheral blood TAZ after operation in the operation group had no significant change compared with the control group (P＞0.05).The level of peripheral blood TAZ in the recurrent group was significantly higher than preoperative level of operation group(P＜0.05).The expression of tumor tissue TAZ was significantly higher than that of normal mucosal tissue(P＜0.05);the expression of tumor tissue TAZ in the recurrent group was higher than that in the operation group(P＜0.05).The tumor tissue TAZ level in the operation group and recurrent group had obvious correlation with peripheral blood TAZ level(P＜0.05).Conclusion Monitoring the TAZ level has obvious reference value for predicting colorectal cancer staging and timely understanding tumor recurrence.

Relationship of TAZ expression between peripheral blood and tumor tissue in patients with colorectal cancer and its clinical significance / 重庆医学

Objective To investigate the expression of TAZ in peripheral blood and tumor tissue of the patients with colorectal cancer and to analyze their relationship and clinical significance.Methods Eighty-five cases of pathologically verified colorectal cancer treated by radical operation were selected as the operation group,and 35 cases with postoperative relapse served as the recurrent group.Meanwhile 50 age and sex-matched healthy persons were selected as the control group.The immunohistochemical(SP)technique was adopted to detect the expression of TAZ in tumor tissues and expression of TAZ in peripheral blood was tested with enzyme linked immunosorbent assay(ELISA).Results The level of preoperative TAZ in peripheral blood of the operation group was significantly higher than that of the control group(P＜0.05).The level of peripheral blood TAZ after operation in the operation group had no significant change compared with the control group (P＞0.05).The level of peripheral blood TAZ in the recurrent group was significantly higher than preoperative level of operation group(P＜0.05).The expression of tumor tissue TAZ was significantly higher than that of normal mucosal tissue(P＜0.05);the expression of tumor tissue TAZ in the recurrent group was higher than that in the operation group(P＜0.05).The tumor tissue TAZ level in the operation group and recurrent group had obvious correlation with peripheral blood TAZ level(P＜0.05).Conclusion Monitoring the TAZ level has obvious reference value for predicting colorectal cancer staging and timely understanding tumor recurrence.

Role of TAZ in Lysophosphatidic Acid-Induced Migration and Proliferation of Human Adipose-Derived Mesenchymal Stem Cells

Transcriptional co-activator with a PDZ-binding motif (TAZ) is an important factor in lysophosphatidic acid (LPA)-induced promotion of migration and proliferation of human mesenchymal stem cells (MSCs). The expression of TAZ significantly increased at 6 h after LPA treatment, and TAZ knockdown inhibited the LPA-induced migration and proliferation of MSCs. In addition, embryonic fibroblasts from TAZ knockout mice exhibited the reduction in LPA-induced migration and proliferation. The LPA1 receptor inhibitor Ki16425 blocked LPA responses in MSCs. Although TAZ knockdown or knockout did not reduce LPA-induced phosphorylation of ERK and AKT, the MEK inhibitor U0126 or the ROCK inhibitor Y27632 blocked LPA-induced TAZ expression along with the reduction in the proliferation and migration of MSCs. Our data suggest that TAZ is an important mediator of LPA signaling in MSCs in the downstream of MEK and ROCK signaling.

Role of TAZ in Lysophosphatidic Acid-Induced Migration and Proliferation of Human Adipose-Derived Mesenchymal Stem Cells

Transcriptional co-activator with a PDZ-binding motif (TAZ) is an important factor in lysophosphatidic acid (LPA)-induced promotion of migration and proliferation of human mesenchymal stem cells (MSCs). The expression of TAZ significantly increased at 6 h after LPA treatment, and TAZ knockdown inhibited the LPA-induced migration and proliferation of MSCs. In addition, embryonic fibroblasts from TAZ knockout mice exhibited the reduction in LPA-induced migration and proliferation. The LPA1 receptor inhibitor Ki16425 blocked LPA responses in MSCs. Although TAZ knockdown or knockout did not reduce LPA-induced phosphorylation of ERK and AKT, the MEK inhibitor U0126 or the ROCK inhibitor Y27632 blocked LPA-induced TAZ expression along with the reduction in the proliferation and migration of MSCs. Our data suggest that TAZ is an important mediator of LPA signaling in MSCs in the downstream of MEK and ROCK signaling.

Regulation of TAZ in cancer

TAZ, a transcriptional coactivator with PDZ-binding motif, is encoded by WWTR1 gene (WW domain containing transcription regulator 1). TAZ is tightly regulated in the hippo pathway-dependent and -independent manner in response to a wide range of extracellular and intrinsic signals, including cell density, cell polarity, F-actin related mechanical stress, ligands of G protein-coupled receptors (GPCRs), cellular energy status, hypoxia and osmotic stress. Besides its role in normal tissue development, TAZ plays critical roles in cell proliferation, differentiation, apoptosis, migration, invasion, epithelial-mesenchymal transition (EMT), and stemness in multiple human cancers. We discuss here the regulators and regulation of TAZ. We also highlight the tumorigenic roles of TAZ and its potential therapeutic impact in human cancers.

Identification of a Novel TAZ Gene Mutation in a Family With X-Linked Dilated Cardiomyopathy Barth Syndrome

Abstract Mutations in the tafazzin (TAZ) gene on chromosome Xq28 are responsible for the Barth syndrome (BTHS) phenotype resulting in a loss of function in the protein tafazzin involved in the transacylation of cardiolipin, an essential mitochondrial phospholipid. TAZ gene was investigated in the proband in our study, who died of dilated cardiomyopathy at 8 months of age, and his family by sequencing to identify the genetic cause of BTHS. Molecular analysis revealed a novel mutation in exon 5 (c.520T>G) of the TAZ gene. This novel mutation c.520T>G, pW174G, was also found in female carriers (mother and grandmother of proband) in the family. Bioinformatic analysis was carried out to examine the effect of mutation in the gene and confirmed the deleterious effect of this single mutation to the protein structure. Protein modeling and 3-dimensional structure of TAZ protein demonstrated the significantly visible changes in mutated protein leading to BTHS phenotype. Prenatal diagnosis in a subsequent pregnancy showed a carrier female, and pregnancy was continued. Child is doing well at 1 year of age.

Clinical and genetic features of Barth syndrome in three patients / 临床儿科杂志

ObjectiveTo explore the clinical presentation, diagnosis, treatment, and outcome of the Barth syndrome (BTHS).MethodsClinical data were collected and analyzed from 3 patients with conifrmed genetic diagnosis of BTHS from June 2013 to October 2014.ResultsAll of the 3 patients were males and two of them were twins. The main clinical manifes-tations of the 3 patients were cardiomyopathy and heart failure, accompanied by different degrees of trabeculations of the left ventricle. Two of them were diagnosed of left ventricular noncompaction (LVNC). All of the 3 patients presented with motor retardation, muscle weakness, growth delay and signiifcantly increased urinary excretion of 3-methylglutaconic acid (3-MGC). One patient was found to have neutropenia. All 3 patients hadTAZ gene mutations which included a novel missense mutation (c.527A>G, p.H176R) detected in the twins and a known nonsense mutation (c.367C>T, p.R123X) identiifed in the other patient. All of the mutations were inherited from their mothers. During the follow-up, the twins died at 7 months old and 7.5 months old respectively. The other patient was still alive.ConclusionBTHS is one of the causes of cardiomyopathy in children. In the male patients who presented with muscle weakness, neutropenia, and increased urinary excretion of 3-MGC, especially in those com-bined with LVNC, BTHS should be screened.