Functions, mechanisms, and therapeutic potential of fetal hemoglobin inducers

胎儿血红蛋白诱导剂的功能、机制和治疗潜力

• 批准号: 10308676
• 负责人: Gerd A Blobel
• 金额: $ 65.29万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-05 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10308676
• 关键词: 5' Untranslated Regions; Accounting; Adult; Animals; Attenuated; Biological Process; Birth; CRISPR screen; CRISPR/Cas technology; Cell Culture Techniques; Cell Line; Cells; Cellular biology; Chromatin; Clone Cells; Codon Nucleotides; Collaborations; Complement; Custom; Data; Defect; Disease; Enzymes; Erythroblasts; Erythrocytes; Erythroid; Erythroid Cells; Fetal Hemoglobin; Foundations; Genes; Genetic Screening; Genetic Transcription; Genetic Translation; Globin; Goals; Hematopoietic stem cells; Hemoglobin; Hemoglobin concentration result; Hemoglobinopathies; Human; Knockout Mice; Laboratories; Libraries; Light; Mass Spectrum Analysis; Measures; Medical; Messenger RNA; Monitor; Mus; Open Reading Frames; PRKR gene; Panthera leo; Patients; Pharmacology; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Pre-Clinical Model; Process; Production; Protein Kinase; Proteins; Proteome; Proteomics; Regulation; Repression; Resolution; Ribosomes; Role; Severity of illness; Sickle Cell Anemia; Sum; Surveys; Testing; Thalassemia; Therapeutic; Therapeutic Index; Tissues; Translations; Western Blotting; Work; base; beta Globin; beta Thalassemia; combinatorial; design; detection limit; druggable target; experimental study; fetal; gamma Globin; improved; improved outcome; insight; mouse model; mutant; novel; restoration; sickling; small molecule; synergism; tool; transcription factor; transcriptome

Abstract Sickle cell disease (SCD) and some types of b-thalassemia that are caused by defects in the adult form of hemoglobin manifest shortly after birth, when the switch from the fetal to the adult form of hemoglobin is complete. Even a partial reversal of this switch is associated with an improved course of these diseases. We employed a newly improved CRISPR-Cas9 platform to carry out a kinase domain-focused genetic screen to identify potentially druggable molecules that repress fetal hemoglobin (HbF) production. This screen uncovered HRI (also known as EIF2AK1), an erythroid-specific protein kinase that regulates protein translation. Depletion of HRI elevates HbF levels in human erythroid cells with few additional perturbations. HRI loss reduces the expression of the major HbF repressor BCL11A, and restoration of BCL11A expression partially restores HbF repression. Moreover, HRI depletion reduces sickling of SCD-derived human erythroid cells in culture. In Aim 1 we will comprehensively dissect HRI function by assessing the transcriptome and proteome of HRI-depleted cells. The goals of Aim 2 are to study the mechanism by which HRI regulates BCL11A, identify additional HRI regulated HbF repressors, and examine the global impact of HRI on protein translational control in primary human erythroid cells. Aim 3 will explore synergies with previously known HbF inducers both using a candidate approach, and by unbiased genetic screens for novel synergies. In Aim 4 we will examine the effects of HRI loss on SCD by generating HRI-deficient humanized SCD mouse models. In sum, these studies explore the role of HRI in human red cell biology and examine HRI as target for pharmacologic HbF induction alone or in combination with mechanistically distinct HbF inducers.

摘要