Endocrine Unit, Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
Proc Natl Acad Sci U S A. 2012 Apr 24;109(17):6638-43. doi: 10.1073/pnas.1117608109. Epub 2012 Apr 10.
Maternal deletion of the NESP55 differentially methylated region (DMR) (delNESP55/ASdel3-4(m), delNAS(m)) from the GNAS locus in humans causes autosomal dominant pseudohypoparathyroidism type Ib (AD-PHP-Ib(delNASm)), a disorder of proximal tubular parathyroid hormone (PTH) resistance associated with loss of maternal GNAS methylation imprints. Mice carrying a similar, maternally inherited deletion of the Nesp55 DMR (ΔNesp55(m)) replicate these Gnas epigenetic abnormalities and show evidence for PTH resistance, yet these mice demonstrate 100% mortality during the early postnatal period. We investigated whether the loss of extralarge αs (XLαs) imprinting and the resultant biallelic expression of XLαs are responsible for the early postnatal lethality in ΔNesp55(m) mice. First, we found that ΔNesp55(m) mice are hypoglycemic and have reduced stomach-to-body weight ratio. We then generated mice having the same epigenetic abnormalities as the ΔNesp55(m) mice but with normalized XLαs expression due to the paternal disruption of the exon giving rise to this Gnas product. These mice (ΔNesp55(m)/Gnasxl(m+/p-)) showed nearly 100% survival up to postnatal day 10, and a substantial number of them lived to adulthood. The hypoglycemia and reduced stomach-to-body weight ratio observed in 2-d-old ΔNesp55(m) mice were rescued in the ΔNesp55(m)/Gnasxl(m+/p-) mice. Surviving double-mutant animals had significantly reduced Gαs mRNA levels and showed hypocalcemia, hyperphosphatemia, and elevated PTH levels, thus providing a viable model of human AD-PHP-Ib. Our findings show that the hypoglycemia and early postnatal lethality caused by the maternal deletion of the Nesp55 DMR result from biallelic XLαs expression. The double-mutant mice will help elucidate the pathophysiological mechanisms underlying AD-PHP-Ib.
人类中 NESP55 差异甲基化区域(DMR)(delNESP55/ASdel3-4(m),delNAS(m))的母体缺失会导致常染色体显性假性甲状旁腺功能减退症 1b 型(AD-PHP-Ib(delNASm)),这是一种近端肾小管甲状旁腺激素(PTH)抵抗的疾病,与母体 GNAS 甲基化印记的丧失有关。携带类似的、母体遗传的 Nesp55 DMR 缺失(ΔNesp55(m))的小鼠复制了这些 Gnas 表观遗传异常,并表现出 PTH 抵抗的证据,但这些小鼠在出生后的早期阶段有 100%的死亡率。我们研究了是否失去特大αs(XLαs)印迹以及由此导致的 XLαs 的双等位基因表达是导致 ΔNesp55(m) 小鼠出生后早期死亡的原因。首先,我们发现 ΔNesp55(m) 小鼠低血糖,胃体比降低。然后,我们生成了具有与 ΔNesp55(m) 小鼠相同的表观遗传异常的小鼠,但由于父本对产生此 Gnas 产物的外显子的破坏,导致 XLαs 的表达正常化。这些小鼠(ΔNesp55(m)/Gnasxl(m+/p-))在出生后第 10 天之前几乎 100%存活,其中相当一部分活到成年。在 2 天大的 ΔNesp55(m) 小鼠中观察到的低血糖和胃体比降低在 ΔNesp55(m)/Gnasxl(m+/p-) 小鼠中得到了挽救。幸存的双突变动物的 Gαs mRNA 水平显著降低,并表现出低钙血症、高磷血症和 PTH 水平升高,从而提供了人类 AD-PHP-Ib 的可行模型。我们的研究结果表明,由 Nesp55 DMR 的母体缺失引起的低血糖和出生后早期死亡是由 XLαs 的双等位基因表达引起的。双突变小鼠将有助于阐明 AD-PHP-Ib 的病理生理机制。