Sharma Pushkal, Kim Colin Y, Keys Heather R, Imada Shinya, Joseph Alex B, Ferro Luke, Kunchok Tenzin, Anderson Rachel, Yilmaz Omer, Weng Jing-Ke, Jain Ankur
Whitehead Institute of Biomedical Research, Cambridge, MA, USA.
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
bioRxiv. 2024 Nov 17:2024.08.24.609500. doi: 10.1101/2024.08.24.609500.
Polyamines are abundant and evolutionarily conserved metabolites that are essential for life. Dietary polyamine supplementation extends life-span and health-span. Dysregulation of polyamine homeostasis is linked to Parkinson's disease and cancer, driving interest in therapeutically targeting this pathway. However, measuring cellular polyamine levels, which vary across cell types and states, remains challenging. We introduce a first-in-class genetically encoded polyamine reporter for real-time measurement of polyamine concentrations in single living cells. This reporter utilizes the polyamine-responsive ribosomal frameshift motif from the OAZ1 gene. We demonstrate broad applicability of this approach and reveal dynamic changes in polyamine levels in response to genetic and pharmacological perturbations. Using this reporter, we conducted a genome-wide CRISPR screen and uncovered an unexpected link between mitochondrial respiration and polyamine import, which are both risk factors for Parkinson's disease. By offering a new lens to examine polyamine biology, this reporter may advance our understanding of these ubiquitous metabolites and accelerate therapy development.
多胺是丰富且在进化上保守的代谢产物，对生命至关重要。饮食中补充多胺可延长寿命和健康期。多胺稳态失调与帕金森病和癌症相关，这引发了对以该途径为治疗靶点的兴趣。然而，测量细胞内多胺水平仍具有挑战性，因为其在不同细胞类型和状态下会有所不同。我们引入了一种一流的基因编码多胺报告基因，用于实时测量单个活细胞中的多胺浓度。该报告基因利用了来自OAZ1基因的多胺响应核糖体移码基序。我们证明了这种方法具有广泛的适用性，并揭示了多胺水平对基因和药物扰动的动态变化。利用该报告基因，我们进行了全基因组CRISPR筛选，发现了线粒体呼吸和多胺导入之间意想不到的联系，而这两者都是帕金森病的风险因素。通过提供一个新的视角来研究多胺生物学，该报告基因可能会增进我们对这些普遍存在的代谢产物的理解，并加速治疗方法的开发。
