PHOSPHOFRUCTOKINASE (PFK) DEFICIENCY

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。除了红细胞丙酮酸激酶缺陷，我们还建立了一个殖民地的M型PFK缺陷的狗。这些动物经历慢性溶血性贫血，伴有碱诱导的溶血危象和劳力性肌病。该缺陷是由过早的终止密码子引起的，导致PFK蛋白的截短（α螺旋的损失）和不稳定性。红细胞中PFK活性的缺乏导致ATP和2，3-二磷酸甘油酯产生受损以及红细胞存活减少。我们已经建立了初步的合作，以研究潜在的治疗干预措施。最近的研究表明，氨基糖苷类药物的给药可能允许通过异常终止密码子的阅读，从而允许正常蛋白质的产生。不幸的是，氨基糖苷类药物具有肾毒性，特别是当使用较长时间时，因此它们不能安全地以可能需要的剂量给药。然而，我们最近接触了一家公司，该公司已经生产了一种类似物，其在狗中的安全性已经得到证明，并计划对囊性纤维化儿童进行I期临床试验。在肌细胞培养物中进行体外试验后，我们计划在PFK缺陷犬中进行初步研究，并监测给药后红细胞中的溶血程度、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，威斯康星州医学院的生物化学副教授Richard Sabine博士与我们接洽，合作进行一项可行性研究，研究使用腺苷产品改善PFK和PK缺陷犬的ATP和DPG产量，然后将这种治疗方法用于患有这些疾病和其他影响ATP合成的疾病的人类患者。

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.