Hanna Joseph, Touahri Yacine, Pak Alissa, Belfiore Lauren, van Oosten Edwin, David Luke Ajay, Han Sisu, Ilnytskyy Yaroslav, Kovalchuk Igor, Kurrasch Deborah, Okawa Satoshi, Del Sol Antonio, Screaton Robert A, Aubert Isabelle, Schuurmans Carol
Biological Sciences, Sunnybrook Research Institute, Toronto, Canada.
Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada.
Elife. 2025 Jun 17;13:RP100604. doi: 10.7554/eLife.100604.
Metabolic pathways are remodeled in response to energy and other homeostatic demands and are dynamically regulated during embryonic development, suggesting a role in guiding cellular differentiation. Here, we show that glycolytic flux is required and sufficient to bias multipotent retinal progenitor cells (RPCs) to acquire a rod photoreceptor fate in the murine retina. In RPC-specific conditional knockout (cKO) and RPC-specific conditional gain-of-function of dominant active PFKFB3 (cytoPFKFB3) mice, glycolytic gene expression and activity are elevated, correlating with precocious rod photoreceptor differentiation and outer segment (OS) maturation. Conversely, glycolytic inhibition in retinal explants suppresses RPC proliferation and photoreceptor differentiation, achieved either with 2-deoxy-D-glucose, a competitive inhibitor of glucose metabolism, by lowering media pH, which disables PKM2, a rate-limiting enzyme, or by inhibiting lactate/H symporters, which lowers intracellular pH. Mechanistically, we show that Wnt signaling, the top-upregulated pathway in cKO retinas, is a glycolysis-dependent pathway. Pharmacological and genetic perturbation of Wnt signaling by knocking-out , encoding β-catenin, phenocopies glycolytic inhibition, suppressing RPC proliferation, photoreceptor differentiation, and OS maturation. Thus, developmental rewiring of glycolytic flux modulates Wnt signaling to drive rod photoreceptor differentiation and maturation, an instructive role that may be exploited therapeutically for cell replacement strategies.
代谢途径会根据能量和其他稳态需求进行重塑，并在胚胎发育过程中受到动态调节，这表明其在引导细胞分化中发挥作用。在此，我们表明糖酵解通量对于使多能视网膜祖细胞（RPCs）偏向于在小鼠视网膜中获得视杆光感受器命运是必需且充分的。在RPC特异性条件性敲除（cKO）小鼠以及RPC特异性条件性过表达显性活性PFKFB3（cytoPFKFB3）的小鼠中，糖酵解基因的表达和活性升高，这与视杆光感受器的早熟分化和外段（OS）成熟相关。相反，视网膜外植体中的糖酵解抑制会抑制RPC增殖和光感受器分化，这可以通过以下方式实现：使用葡萄糖代谢的竞争性抑制剂2-脱氧-D-葡萄糖、通过降低培养基pH值（这会使限速酶PKM2失活）或通过抑制乳酸/氢离子同向转运体（这会降低细胞内pH值）。从机制上讲，我们表明Wnt信号通路是cKO视网膜中上调最明显的通路，是一条糖酵解依赖性通路。通过敲除编码β-连环蛋白的基因对Wnt信号通路进行药理学和遗传学干扰，模拟了糖酵解抑制，抑制了RPC增殖、光感受器分化和OS成熟。因此，糖酵解通量的发育性重编程调节Wnt信号通路以驱动视杆光感受器的分化和成熟，这一指导作用可能在细胞替代策略的治疗中得到利用。
