Brandt Annely, Petrovsky Roman, Kriebel Maria, Großhans Jörg
Bieneninstitut Kirchhain, 35274 Kirchhain, Germany.
Department of Biology, Philipps University, Karl-von-Frisch-Straße 8, 35043 Marburg, Germany.
J Dev Biol. 2023 Oct 29;11(4):40. doi: 10.3390/jdb11040040.
The presence of farnesylated proteins at the inner nuclear membrane (INM), such as the Lamins or Kugelkern in , leads to specific changes in the nuclear morphology and accelerated ageing on the organismal level reminiscent of the Hutchinson-Gilford progeria syndrome (HGPS). Farnesyl transferase inhibitors (FTIs) can suppress the phenotypes of the nuclear morphology in cultured fibroblasts from HGPS patients and cultured cells overexpressing farnesylated INM proteins. Similarly, FTIs have been reported to suppress the shortened lifespan in model organisms. Here, we report an experimental system combining cell culture and flies for testing the activity of substances on the HGPS-like nuclear morphology and lifespan, with FTIs as an experimental example. Consistent with previous reports, we show that FTIs were able to ameliorate the nuclear phenotypes induced by the farnesylated nuclear proteins Progerin, Kugelkern, or truncated Lamin B in cultured cells. The subsequent validation in lifespan assays demonstrated the applicability of the experimental system: treating adult with the FTI ABT-100 reversed the nuclear phenotypes and extended the lifespan of experimentally induced short-lived flies. Since -expressing flies have a significantly shorter average lifespan, half the time is needed for testing substances in the lifespan assay.
在内核膜（INM）上存在法尼基化蛋白，如核纤层蛋白或中的Kugelkern，会导致核形态发生特定变化，并在机体水平上加速衰老，这让人联想到哈钦森 - 吉尔福德早衰综合征（HGPS）。法尼基转移酶抑制剂（FTIs）可以抑制HGPS患者培养的成纤维细胞以及过表达法尼基化INM蛋白的培养细胞中的核形态表型。同样，据报道FTIs可以抑制模式生物缩短的寿命。在这里，我们报告了一个结合细胞培养和果蝇的实验系统，用于测试物质对HGPS样核形态和寿命的活性，并以FTIs作为实验示例。与之前的报道一致，我们表明FTIs能够改善培养细胞中法尼基化核蛋白早老素、Kugelkern或截短的核纤层蛋白B诱导的核表型。随后在果蝇寿命测定中的验证证明了该实验系统的适用性：用FTI ABT - 100处理成年果蝇可逆转核表型并延长实验诱导的短寿命果蝇的寿命。由于表达的果蝇平均寿命明显较短，因此在寿命测定中测试物质所需的时间减半。
