作为一种重要的转录激活剂，PDX1 在胰腺发育和 β 细胞功能中发挥着至关重要的作用。 PDX1 基因突变可能导致 4 型青少年发病型糖尿病 (MODY4) 和新生儿糖尿病。然而，由于临床样本的缺乏以及人类和现有动物模型之间胰腺结构和基因组组成的显着差异，MODY4 的精确机制仍然难以捉摸。在这项研究中，利用 CRISPR/Cas9 技术培育了三只 PDX1 突变食蟹猴，它们均在产后不久死亡，表现出胰腺发育不全。值得注意的是，一只三等位基因 PDX1 突变型食蟹猴（称为 M4）发育出胰腺，而两只单等位基因 PDX1 突变型食蟹猴则没有显示出胰腺形成的解剖学证据。 M4 胰腺的 RNA 测序揭示了内分泌和外分泌功能的显着分子变化，表明发育迟缓和 PDX1 单倍体不足。通过培养的 PDX1 突变胰岛类器官证实了 M4 胰腺中 m6A 甲基化的显着变化。值得注意的是，m6A 调节剂 METTL3 的过度表达恢复了杂合 PDX1 突变胰岛类器官的功能。这项研究强调了 m6A 甲基化修饰在 MODY4 进展中的新作用，并为临床前研究提供了有价值的分子见解。

As an essential transcriptional activator, PDX1 plays a crucial role in pancreatic development and β-cell function. Mutations in the PDX1 gene may lead to type 4 maturity-onset diabetes of the young (MODY4) and neonatal diabetes mellitus. However, the precise mechanisms underlying MODY4 remain elusive due to the paucity of clinical samples and pronounced differences in pancreatic architecture and genomic composition between humans and existing animal models. In this study, three PDX1-mutant cynomolgus macaques were generated using CRISPR/Cas9 technology, all of which succumbed shortly postpartum, exhibiting pancreatic agenesis. Notably, one tri-allelic PDX1-mutant cynomolgus macaque (designated as M4) developed a pancreas, whereas the two mono-allelic PDX1-mutant cynomolgus macaques displayed no anatomical evidence of pancreatic formation. RNA sequencing of the M4 pancreas revealed substantial molecular changes in both endocrine and exocrine functions, indicating developmental delay and PDX1 haploinsufficiency. A marked change in m6A methylation was identified in the M4 pancreas, confirmed through cultured PDX1-mutant islet organoids. Notably, overexpression of the m6A modulator METTL3 restored function in heterozygous PDX1-mutant islet organoids. This study highlights a novel role of m6A methylation modification in the progression of MODY4 and provides valuable molecular insights for preclinical research.