Waite Eric L, Tigue Mark, Yu Ming, Lahori Deeksha, Kelly Kai, May Catherine Lee, Naji Ali, Roman Jeffrey, Doliba Nicolai, Avrahami Dana, Nguyen-Ngoc Kim-Vy, Sander Maike, Glaser Benjamin, Kaestner Klaus H
Institute of Diabetes, Obesity, and Metabolism, Perelman School of Medicine, The University of Pennsylvania, Philadelphia, PA.
Department of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA.
Diabetes. 2025 Mar 1;74(3):332-342. doi: 10.2337/db23-0887.
The gold standard for assessing the function of human islets or β-like cells derived from stem cells involves their engraftment under the kidney capsule of hyperglycemic, immunodeficient mice. Current models, such as streptozotocin treatment of severely immunodeficient mice or the NRG-Akita strain, are limited due to unstable and variable hyperglycemia and/or high morbidity. To address these limitations, we developed the IsletTester mouse via CRISPR/Cas9-mediated gene editing of glucokinase (Gck), the glucose sensor of the β-cells, directly in NSG zygotes. IsletTester mice are heterozygous for an Arg345→stop mutation in Gck and present with stable random hyperglycemia (∼250 mg/dL [14 mmol/L]), normal lifespan, and fertility. We demonstrate the utility of this model through functional engraftment of both human islets and human embryonic stem cell-derived β-like cells. The IsletTester mouse will enable the study of human islet biology over time and under different physiological conditions and can provide a useful preclinical platform to determine the functionality of stem cell-derived islet products.
Current mouse models for assessing islet function in vivo are limited due to unstable and variable hyperglycemia and/or high morbidity. We derived the IsletTester mouse to address these limitations. Leveraging a previously characterized glucokinase mutation and CRISPR/Cas9 technology, we successfully developed a moderately hyperglycemic and immunodeficient mouse model for the in vivo assessment of islet function. Our IsletTester mouse has stable, moderate hyperglycemia that can be corrected with primary human islets or stem cell-derived insulin-producing cells. The IsletTester mouse provides a reliable, easy-to-use platform for the preclinical assessment of stem cell-derived islet products or islet-targeted drugs.
评估人胰岛或源自干细胞的β样细胞功能的金标准是将它们移植到高血糖、免疫缺陷小鼠的肾包膜下。当前的模型，如用链脲佐菌素处理严重免疫缺陷小鼠或NRG-Akita品系，由于高血糖不稳定且变化不定和/或发病率高而受到限制。为了解决这些局限性，我们通过对β细胞的葡萄糖传感器葡萄糖激酶（Gck）进行CRISPR/Cas9介导的基因编辑，直接在NSG受精卵中培育出了胰岛测试小鼠。胰岛测试小鼠对于Gck中的Arg345→终止突变是杂合的，表现出稳定的随机高血糖（约250mg/dL[14mmol/L]）、正常寿命和生育能力。我们通过人胰岛和人胚胎干细胞衍生的β样细胞的功能移植证明了该模型的实用性。胰岛测试小鼠将能够随着时间推移并在不同生理条件下研究人胰岛生物学，并可为确定干细胞衍生的胰岛产品的功能提供一个有用的临床前平台。
当前用于体内评估胰岛功能的小鼠模型由于高血糖不稳定且变化不定和/或发病率高而受到限制。我们培育出胰岛测试小鼠以解决这些局限性。利用先前表征的葡萄糖激酶突变和CRISPR/Cas9技术，我们成功开发了一种中度高血糖和免疫缺陷的小鼠模型，用于体内评估胰岛功能。我们的胰岛测试小鼠具有稳定的中度高血糖，可用原代人胰岛或干细胞衍生的胰岛素生成细胞纠正。胰岛测试小鼠为干细胞衍生的胰岛产品或靶向胰岛的药物的临床前评估提供了一个可靠、易用的平台。
