Santillan Donna A, Santillan Mark K, Hunter Stephen K
Department of Obstetrics and Gynecology, University of Iowa Hospitals and Clinics, Iowa City, IA 52242, USA.
Am J Obstet Gynecol. 2009 Sep;201(3):289.e1-6. doi: 10.1016/j.ajog.2009.05.035. Epub 2009 Jul 24.
The objective of this work was to determine whether cells overexpressing phenylalanine (Phe) hydroxylase (PAH) can significantly reduce Phe in vitro for potential use as a therapy for preventing maternal phenylketonuria.
Human 293T and WRL68 cell lines were transiently and stably transfected to overexpress PAH. Cells were encapsulated within microspheres of sodium alginate. Timed measurements of Phe in media were performed using tandem mass spectrometry.
Both nonencapsulated and encapsulated transiently transfected cells overexpressing PAH significantly reduced the Phe concentration in media by approximately 50% in comparison to mock-transfected cells. Cell line clones stably expressing PAH significantly decreased the Phe concentration in the media by up to 85% compared with media alone.
Both unencapsulated and encapsulated cells overexpressing PAH significantly reduce Phe in vitro. Studies using phenylketonuria model mice will be important in determining the ability of our therapy to prevent the teratogenic effects of elevated maternal Phe in maternal phenylketonuria.
本研究旨在确定过表达苯丙氨酸（Phe）羟化酶（PAH）的细胞是否能在体外显著降低苯丙氨酸水平，从而有可能作为预防母体苯丙酮尿症的一种治疗方法。
对人293T和WRL68细胞系进行瞬时和稳定转染，使其过表达PAH。将细胞封装在海藻酸钠微球中。使用串联质谱法对培养基中的苯丙氨酸进行定时测量。
与mock转染细胞相比，过表达PAH的未封装和封装的瞬时转染细胞均使培养基中的苯丙氨酸浓度显著降低了约50%。与单独的培养基相比，稳定表达PAH的细胞系克隆使培养基中的苯丙氨酸浓度显著降低了高达85%。
过表达PAH的未封装和封装细胞均能在体外显著降低苯丙氨酸水平。使用苯丙酮尿症模型小鼠进行的研究对于确定我们的治疗方法预防母体苯丙酮尿症中母体苯丙氨酸升高的致畸作用的能力至关重要。
