Choe Donghui, Palsson Bernhard O
Shu Chien-Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA.
Department of Pediatrics, University of California San Diego, La Jolla, CA, USA.
Nat Biomed Eng. 2025 Apr 30. doi: 10.1038/s41551-025-01391-y.
Advances in high-throughput sequencing have enabled the identification of genetic variations associated with human disease. However, deciphering the functional significance of these variations remains challenging. Here we propose an alternative approach that uses humanized Escherichia coli to study human genetic enzymopathies and to screen candidate drug effects on metabolic targets. By replacing selected E. coli metabolic enzymes with their human orthologues and their sequence variants, we demonstrate that the growth rate of E. coli reflects the in vivo activity of heterologously expressed human enzymes. This approach accurately reflected enzyme activities of known sequence variants, enabling rapid screening of causal sequence variations associated with human diseases. This approach bridges the gap between in vitro assays and cell-based assays. Our findings suggest that the proposed approach using a humanized E. coli strain holds promise for drug discovery, offering a high-throughput and cost-effective platform for identifying new compounds targeting human enzymes. Continued research and innovation in this field have the potential to impact the development and practice of precision medicine.
高通量测序技术的进步使得与人类疾病相关的基因变异得以识别。然而，解读这些变异的功能意义仍然具有挑战性。在此，我们提出一种替代方法，该方法利用人源化大肠杆菌来研究人类遗传酶病，并筛选候选药物对代谢靶点的作用。通过用其人类直系同源物及其序列变体替代选定的大肠杆菌代谢酶，我们证明大肠杆菌的生长速率反映了异源表达的人类酶在体内的活性。这种方法准确反映了已知序列变体的酶活性，能够快速筛选与人类疾病相关的致病序列变异。这种方法弥合了体外试验和基于细胞的试验之间的差距。我们的研究结果表明，所提出的使用人源化大肠杆菌菌株的方法在药物发现方面具有前景，为识别靶向人类酶的新化合物提供了一个高通量且经济高效的平台。该领域持续的研究和创新有可能影响精准医学的发展和实践。
