Pathogenetics of the FOX transcription factor gene cluster on 16q24.1

16q24.1 FOX转录因子基因簇的发病机制

• 批准号: 8460859
• 负责人: PAWEL STANKIEWICZ
• 金额: $ 42.5万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8460859
• 关键词: Address; Adenovirus Vector; Affect; Alleles; Alveolar; Amniotic Fluid; Benefits and Risks; Binding Sites; Biochemical; Biological Assay; Birth; Blood capillaries; Blood-Air Barrier; Boxing; Candidate Disease Gene; Cardiac; ChIP-on-chip; Characteristics; Chromatin; Communication; Complex; Congenital Abnormality; Copy Number Polymorphism; Data; Defect; Development; Diffuse; Disease; Dysplasia; Embryo; Endothelium; Epithelial Cells; Financial compensation; Gene Cluster; Gene Expression Regulation; Gene Targeting; Gene Transfer; Genes; Genitourinary system; Genomic Imprinting; Genomics; Hemorrhage; Hospitals; Human; Hypoplastic Left Heart Syndrome; Infant; Insulator Elements; Intensive Care; Investigational Therapies; Knowledge; Life; Lung; Lung diseases; Measures; Mesenchyme; Methods; Molecular; Molecular Conformation; Mus; Mutation; Neonatal; Newborn Infant; Organ; Pathogenesis; Pathway interactions; Patients; Pattern; Peripheral; Phase; Phenotype; Physiological; Point Mutation; Positioning Attribute; Pregnancy; Protein Binding; Proteins; Pulmonary Hypertension; Pulmonary veins; Regulation; Regulatory Element; Reporter; Respiratory distress; Risk; Rodent; Route; Safety; Structure of parenchyma of lung; Symptoms; Techniques; Testing; Therapeutic; Time; Tissues; Transcription factor genes; Translating; Vascularization; Viral Vector; Weaning; base; capillary; design; developmental disease; dosage; forkhead protein; gastrointestinal; gene therapy; helper-dependent adenoviral vector; in utero; loss of function; lung development; microdeletion; nonhuman primate; postnatal; prevent; public health relevance; screening; segregation; transcription factor; vasculogenesis

DESCRIPTION (provided by applicant): Alveolar capillary dysplasia with misalignment of pulmonary veins (ACD/MPV) is a lethal neonatal diffuse developmental disorder of the lungs, which is commonly associated with multiple congenital anomalies involving the cardiac, gastrointestinal, and genitourinary systems. Infants affected with ACD/MPV develop severe respiratory distress with pulmonary hypertension within the first few days of life and despite intensive care they usually die soon thereafter. Recently, we have found that haploinsufficiency due to point mutations or genomic deletions of the transcription factor Forkhead Box F1 (FOXF1) on 16q24.1 results in ACD/MPV and a broad spectrum of congenital malformations. In addition, we have identified two distinct microdeletions upstream of FOXF1, implicating a position effect in the pathogenesis of the disease. Pleiotropic effects encountered in FOXF1 microdeletions, such as hypoplastic left heart syndrome and gastrointestinal atresias, may be due to haploinsufficiency for the neighboring genes, FOXC2 and FOXL1, both part of the FOX cluster at 16q24.1. Heterozygous Foxf1 mice die from pulmonary hemorrhage with severe defects in lung alveolarization and vasculogenesis along with other organ anomalies, although they do not completely recapitulate ACD/MPV in humans. The expression of the Foxf1 gene during development suggests an intriguing pattern of gene regulation. We hypothesize that this complex regulation of Foxf1 may be due to both position effects and genomic imprinting in a tissue- and time-specific manner; ACD/MPV can also be caused by disruption of other gene(s) or FOXF1 regulatory elements; and the lung defect in Foxf1 mice can be prevented perinatally by increasing the dosage of the Foxf1 protein in the capillary endothelium and surrounding mesenchyme. We have designed three aims to test these hypotheses. In aim 1, we will dissect gene regulation of FOXF1 in two ways. First, regulatory elements that may be important to the expression of the FOX gene cluster or to FOXF1 specific expression will be identified and tested using reporter assays, ChIP-on-chip, and chromatin conformation capture (3C) techniques. Second, we will analyze the segregation and allele-specific expression of Foxf1 in mice. In aim 2, we will use the knowledge gained from our studies in aim 1 to screen for point mutations and copy-number variations in the regulatory elements identified upstream or downstream to FOXF1 and in the ACD/MPV candidate genes. Finally, in aim 3, we will explore therapeutic options by use of adenoviral vector-based Foxf1 gene transfer in peripheral murine lungs. ACD/MPV is a lethal disorder and there is no available treatment at the present time. We believe that a gene therapy approach using viral vectors may have the potential to correct the lethal phenotype of ACD/MPV patients by reversing the abnormal formation of the lethal capillary defect. Moreover, the risks related to this experimental therapy may be justified from a risk: benefit standpoint and have potential to be translated in the hospital setting.

描述（由申请人提供）：肺泡毛细血管发育不良伴肺静脉错位（ACD/MPV）是一种致命的新生儿弥漫性肺部发育障碍，通常与涉及心脏、胃肠道和泌尿生殖系统的多种先天性异常有关。感染 ACD/MPV 的婴儿在出生后的最初几天内会出现严重的呼吸窘迫和肺动脉高压，尽管经过重症监护，他们通常很快就会死亡。最近，我们发现由于 16q24.1 上转录因子 Forkhead Box F1 (FOXF1) 的点突变或基因组缺失导致的单倍体不足，导致 ACD/MPV 和广泛的先天畸形。此外，我们还发现了 FOXF1 上游的两个不同的微缺失，暗示了该疾病发病机制中的位置效应。 FOXF1 微缺失中遇到的多效性效应，例如左心发育不全综合征和胃肠道闭锁，可能是由于邻近基因 FOXC2 和 FOXL1（均为 16q24.1 处 FOX 簇的一部分）的单倍体不足所致。杂合性 Foxf1 小鼠死于肺出血，并伴有肺泡化和血管生成的严重缺陷以及其他器官异常，尽管它们不能完全重现人类的 ACD/MPV。 Foxf1 基因在发育过程中的表达表明了一种有趣的基因调控模式。我们假设 Foxf1 的这种复杂调控可能是由于位置效应和组织和时间特异性方式的基因组印记所致。 ACD/MPV 也可能是由其他基因或 FOXF1 调控元件破坏引起的；围产期增加Foxf1蛋白在毛细血管内皮和周围间质中的剂量可以预防Foxf1小鼠的肺缺陷。我们设计了三个目标来检验这些假设。在目标 1 中，我们将以两种方式剖析 FOXF1 的基因调控。首先，将使用报告基因检测、ChIP-on-chip 和染色质构象捕获 (3C) 技术来鉴定和测试对 FOX 基因簇表达或 FOXF1 特异性表达可能重要的调控元件。其次，我们将分析 Foxf1 在小鼠中的分离和等位基因特异性表达。在目标 2 中，我们将利用从目标 1 的研究中获得的知识来筛选 FOXF1 上游或下游以及 ACD/MPV 候选基因中鉴定的调控元件中的点突变和拷贝数变异。最后，在目标 3 中，我们将通过在小鼠外周肺中使用基于腺病毒载体的 Foxf1 基因转移来探索治疗方案。 ACD/MPV 是一种致命性疾病，目前尚无可用的治疗方法。我们相信，使用病毒载体的基因治疗方法可能有潜力通过逆转致死性毛细血管缺陷的异常形成来纠正 ACD/MPV 患者的致死表型。此外，与这种实验性疗法相关的风险可能从风险：效益的角度来看是合理的，并且有可能在医院环境中转化。

项目成果

Molecular and clinical analyses of 16q24.1 duplications involving FOXF1 identify an evolutionarily unstable large minisatellite.

• DOI: 10.1186/s12881-014-0128-z
• 发表时间: 2014-12-04
• 期刊: BMC medical genetics
• 作者: Dharmadhikari AV;Gambin T;Szafranski P;Cao W;Probst FJ;Jin W;Fang P;Gogolewski K;Gambin A;George-Abraham JK;Golla S;Boidein F;Duban-Bedu B;Delobel B;Andrieux J;Becker K;Holinski-Feder E;Cheung SW;Stankiewicz P
• 通讯作者: Stankiewicz P

Small noncoding differentially methylated copy-number variants, including lncRNA genes, cause a lethal lung developmental disorder.

• DOI: 10.1101/gr.141887.112
• 发表时间: 2013-01
• 期刊: Genome research
• 影响因子: 7
• 作者: Szafranski P;Dharmadhikari AV;Brosens E;Gurha P;Kolodziejska KE;Zhishuo O;Dittwald P;Majewski T;Mohan KN;Chen B;Person RE;Tibboel D;de Klein A;Pinner J;Chopra M;Malcolm G;Peters G;Arbuckle S;Guiang SF 3rd;Hustead VA;Jessurun J;Hirsch R;Witte DP;Maystadt I;Sebire N;Fisher R;Langston C;Sen P;Stankiewicz P
• 通讯作者: Stankiewicz P

Variable phenotypic presentation of a novel FOXF1 missense mutation in a single family.

单一家族中新型 FOXF1 错义突变的可变表型表现。

• DOI: 10.1002/ppul.23425
• 发表时间: 2016
• 期刊: Pediatric pulmonology
• 影响因子: 3.1
• 作者: Reiter,Joel;Szafranski,Przemyslaw;Breuer,Oded;Perles,Zeev;Dagan,Tamir;Stankiewicz,Paweł;Kerem,Eitan
• 通讯作者: Kerem,Eitan

16q24.1 microdeletion in a premature newborn: usefulness of array-based comparative genomic hybridization in persistent pulmonary hypertension of the newborn.

早产儿 16q24.1 微缺失：基于阵列的比较基因组杂交在新生儿持续性肺动脉高压中的有用性。

• DOI: 10.1097/pcc.0b013e3182192c96
• 发表时间: 2011
• 期刊: Pediatric critical care medicine : a journal of the Society of Critical Care Medicine and the World Federation of Pediatric Intensive and Critical Care Societies
• 作者: Zufferey,Flore;Martinet,Danielle;Osterheld,Maria-Chiara;Niel-Butschi,Florence;Giannoni,Eric;Schmutz,NathalieBesuchet;Xia,Zhilian;Beckmann,JacquesS;Shaw-Smith,Charles;Stankiewicz,Pawel;Langston,Claire;Fellmann,Florence
• 通讯作者: Fellmann,Florence

Comparative analyses of lung transcriptomes in patients with alveolar capillary dysplasia with misalignment of pulmonary veins and in foxf1 heterozygous knockout mice.

肺泡毛细血管发育不良伴肺静脉错位患者与foxf1杂合基因敲除小鼠肺转录组的比较分析。

• DOI: 10.1371/journal.pone.0094390
• 发表时间: 2014
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Sen,Partha;Dharmadhikari,AvinashV;Majewski,Tadeusz;Mohammad,MahmoudA;Kalin,TanyaV;Zabielska,Joanna;Ren,Xiaomeng;Bray,Molly;Brown,HannahM;Welty,Stephen;Thevananther,Sundararajah;Langston,Claire;Szafranski,Przemyslaw;Justice,M
• 通讯作者: Justice,M