PHOSPHOFRUCTOKINASE (PFK) DEFICIENCY

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。除了红细胞丙酮酸激酶缺乏症外，我们还建立了一个m型PFK缺陷犬群。这些动物经历慢性溶血性贫血，伴有碱性引起的溶血危象和运动性肌病。该缺陷是由过早终止密码子导致截断（α螺旋的丢失）和PFK蛋白的不稳定引起的。红细胞中缺乏PFK活性会导致ATP和2,3-二磷酸甘油酯的产生受损，并降低红细胞的存活率。我们已经建立了初步合作，以调查潜在的治疗干预措施。最近的研究表明，氨基糖苷类的管理可能允许读取通过一个异常的停止密码子，从而允许生产正常的蛋白质。不幸的是，氨基糖苷具有肾毒性，特别是在较长时间使用时，因此不能安全地按可能需要的剂量给药。然而，最近有一家公司与我们接洽，该公司生产了一种类似物，其在狗身上的安全性已经得到证实，并计划对患有囊性纤维化的儿童进行一期临床试验。在肌肉细胞培养的体外测试之后，我们计划在PFK缺乏的狗身上进行一项试点研究，并监测给药后红细胞的溶血程度、红细胞代谢产物、血红蛋白-氧解离曲线、碱性脆性和实际PFK活性。此外，威斯康星医学院生物化学副教授Richard Sabine博士与我们进行了合作，在对患有这些疾病和其他影响ATP合成的疾病的人类患者使用这种治疗方法之前，合作进行一项关于使用腺苷产品改善PFK和PK缺陷犬的ATP和DPG生成的可行性研究。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In addition to the erythrocytic pyruvate kinase deficiency, we have also established a colony of M-type PFK deficient dogs. These animals experience a chronic hemolytic anemia with alkaline induced hemolytic crises and exertional myopathy. The defect is caused by a premature stop codon leading to a truncation (loss of an alpha helix) and instability of the PFK protein. The lack of PFK activity in erythrocytes leads to impaired ATP and 2,3-diphosphoglyceride production and reduced survival of erythrocytes. We have established preliminary collaborations to investigate potential therapeutic interventions. Recent studies have shown that the administration of aminoglycosides may permit the read through an aberrant stop codon thereby permitting the production of the normal protein. Unfortunately, aminoglycosides are nephrotoxic particularly when used for longer periods of time and therefore they cannot safely be administered at the dose likely required. However, we have recently been approached by a company that has produced an analogue for which the safety in dogs has already been documented and a phase I clinical trial for children with cystic fibrosis is planned. Following in vitro testing in muscle cell cultures, we plan to perform a pilot study in the PFK deficient dogs and monitor the degree of hemolysis, red cell metabolites, hemoglobin-oxygen dissociation curve, alkaline fragility, and actual PFK activity in erythrocytes after administration. Furthermore, we have been approached by Richard Sabine, PhD, Associate Professor of Biochemistry from the Medical College of Wisconsin to collaborate in a feasibility study on the use of adenosine products to improve ATP and DPG production in PFK and PK deficient dogs prior to using this therapeutic approach in human patients with these and other diseases affecting ATP synthesis.