QUINARY INTERACTIONS OF HEMOGLOBIN S & DESIGN OF SUPER-ANTISICKLING HEMOGLOBINS

(改编自申请人摘要)脱氧HBs的聚合，尽管 Val-6(Beta)的主要后果是合作参与 其他分子间接触部位。潜在的介绍和介绍能力 在适当的靶细胞中表达基因增加了人们对 镰状细胞病的基因治疗。一种策略涉及到表达 抗镰刀状血红蛋白扰乱患者的侧向接触部位 聚合物。具有超抑制作用的抗镰状血红蛋白的设计 潜力(与短指HBF相比)可以实现临床 低水平表达的好处。初步研究表明， 证明了小鼠的非人类阿尔法链，特别是 猪通过多种物质的互补作用强烈抑制聚合反应 序列差异(关联的多位点扰动)。调查人员 假设有可能通过选择 在物种间，连接最好的多位点序列差异。这个 研究人员已经产生了含有非人α-HBs的嵌合HBs 链以及呈现多个序列的嵌合阿尔法链 差异以及呈现多个序列的嵌合阿尔法链 差异(与人类相比)，旨在绘制这种链接的多站点 微扰。α-珠蛋白链的模数构造(使用 最少两个模块，最多五个模块) 拼接人类和/或非人类阿尔法基因的互补片段 链将被用作产生嵌合α链的主要方法。 猴、马、鼠和猪的a链含有4、18、19和 分别选择了22个序列差异作为初始序列 学习。这些嵌合链将被用来定义内部- 序列差异的四聚体功能互补 引入顺式和反式二聚体，以增加对 聚合反应。嵌合血红蛋白将进行构象测试 可能会改变它们功能的差异。超抑制的阿尔法- 链将与具有序列差异的BetaA链杂交 接收器凹槽区域和/或轴向接触区域以增强 抗镰状血红蛋白的抑制作用。调查人员预计 这种超抑制性血红蛋白的基因结构可能是 欢迎加入用于镰状细胞基因治疗的兵器 疾病。

(Adapted from Applicant's Abstract) Polymerization of deoxy HbS, although a primary consequence of Val-6(Beta), involves a cooperative participation other intermolecular contact sites. The potential ability to introduce and express genes in appropriate target cells has increased the interest in gene therapy of sickle cell disease. One strategy involves the expression of anti-sickling hemoglobins to perturb the lateral contact sites in the polymer. Design of anti-sickling hemoglobins with super inhibitory potential (compared to the pardigmatic HbF) could realize clinical benefits at low levels of expression. The preliminary studies have demonstrated that non-human alpha-chains of mouse and, in particular, of swine strongly inhibit polymerization by complementary effects of many sequence differences (linked multi-site perturbations). The investigators hypothesize that it is possible to optimize this phenomenon by selecting across species, the best linked multi-site sequence differences. The investigators have generated chimeric HbS containing non-human alpha- chains as well as chimeric alpha-chains exhibiting multiple sequence differences The well as chimeric alpha-chains exhibiting multiple sequence differences (compared with human), designed to map such linked multisite perturbations. The modular construction of alpha-globin chains (using a minimum of two and a maximum of five modules) through protease mediated splicing of the complimentary segments of human and/or non-human alpha- chains will be used as the primary approach to generate chimeric a-chains. The a-chains of monkey, horse, mouse, and swine containing 4, 18, 19 and 22 sequence differences, respectively, have been chosen for the initial studies. These chimeric chains will be used to define the intra- tetrameric functional complementarity of the sequence differences introduced into the cis and trans dimers to increase the inhibition of polymerization. Chimeric hemoglobins will be tested for conformational differences that could alter their function. The super-inhibitory alpha- chains will be hybridized with BetaA chains with sequence differences in the acceptor pocket region and/or the axial contact regions to enhance the inhibitory influence of the anti-sickling Hb. The investigators project that gene constructs of such super-inhibitory hemoglobins could be a welcome addition to the armamentarium for the gene therapy of sickle cell disease.