Mechanistic Insight into Serine Flux Regulation through Nanoscale Organization of Glucose and Serine Transporters by Substrate Probe-Based Direct Stochastic Optical Reconstruction Microscopy Imaging

Serine serves as a metabolic nexus in tumors, coordinating one-carbon metabolism, nucleotide synthesis, and redox regulation. While serine transporters (SerTs) are known to be dysregulated in cancer, their functional nanoscale organization remains unresolved due to the limitation of resolution imagi...

Full description

Saved in:
Bibliographic Details
Main Authors: Pengwei Jiang, Hao Hou, Jiaqi Wang, Xumin Wang, Yaqi Wang, Simin Liu, Junling Chen, Hongda Wang, Feng Liang
Format: Article
Language:English
Published: American Association for the Advancement of Science (AAAS) 2025-01-01
Series:Research
Online Access:https://spj.science.org/doi/10.34133/research.0805
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849335765439348736
author Pengwei Jiang
Hao Hou
Jiaqi Wang
Xumin Wang
Yaqi Wang
Simin Liu
Junling Chen
Hongda Wang
Feng Liang
author_facet Pengwei Jiang
Hao Hou
Jiaqi Wang
Xumin Wang
Yaqi Wang
Simin Liu
Junling Chen
Hongda Wang
Feng Liang
author_sort Pengwei Jiang
collection DOAJ
description Serine serves as a metabolic nexus in tumors, coordinating one-carbon metabolism, nucleotide synthesis, and redox regulation. While serine transporters (SerTs) are known to be dysregulated in cancer, their functional nanoscale organization remains unresolved due to the limitation of resolution imaging and available probes. Here, we developed a substrate-based fluorescent probe (Ser-probe) enabling direct stochastic optical reconstruction microscopy of SerTs, revealing malignancy-associated clustering assembly of SerTs that correlates with transport capacity. Compared to MDA-MB-231 cells, MCF7 cells with higher endogenous serine biosynthetic capacity exhibited more pronounced SerT/glucose transporter (GluT) co-clustering, suggesting that their spatial assemblies closely correlate with serine transport and biosynthetic functions in maintaining serine homeostasis. Their cluster morphology and co-assembly were revealed to depend critically on lipid rafts and glycan cross-linking, identifying the key determinants of spatial distribution to enable mechanistic understanding and potential regulation. Glucose deprivation weakened SerT/GluT clustering and their colocalization, which may be caused by their attenuated functional cooperativity in serine homeostasis maintenance under glucose-dependent suppression of serine synthesis. Pharmacological inhibition of phosphoglycerate dehydrogenase (PHGDH) initially enhanced SerT/GluT aggregation and colocalization, but this effect gradually attenuated as doses increased. The strategic combination of a PHGDH inhibitor with glucose restriction or free sialic acid synergistically disrupted SerT/GluT nanoscale organization, amplifying the anti-tumor efficacy of the PHGDH inhibitor and establishing the metabolic plasticity of transporter assemblies as a targetable vulnerability. This work establishes a fundamental link between transporter spatial assembly and tumor serine metabolic reprogramming, providing a new perspective to better understand SerT dysfunction in tumor metabolic reprogramming, offering novel therapeutic avenues for targeting serine metabolism in cancer.
format Article
id doaj-art-84d8448f3db849a384e2b692aacfb807
institution Kabale University
issn 2639-5274
language English
publishDate 2025-01-01
publisher American Association for the Advancement of Science (AAAS)
record_format Article
series Research
spelling doaj-art-84d8448f3db849a384e2b692aacfb8072025-08-20T03:45:10ZengAmerican Association for the Advancement of Science (AAAS)Research2639-52742025-01-01810.34133/research.0805Mechanistic Insight into Serine Flux Regulation through Nanoscale Organization of Glucose and Serine Transporters by Substrate Probe-Based Direct Stochastic Optical Reconstruction Microscopy ImagingPengwei Jiang0Hao Hou1Jiaqi Wang2Xumin Wang3Yaqi Wang4Simin Liu5Junling Chen6Hongda Wang7Feng Liang8The State Key Laboratory of Refractories and Metallurgy, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, Wuhan, Hubei 430081, P. R. China.The State Key Laboratory of Refractories and Metallurgy, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, Wuhan, Hubei 430081, P. R. China.The State Key Laboratory of Refractories and Metallurgy, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, Wuhan, Hubei 430081, P. R. China.The State Key Laboratory of Refractories and Metallurgy, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, Wuhan, Hubei 430081, P. R. China.The State Key Laboratory of Refractories and Metallurgy, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, Wuhan, Hubei 430081, P. R. China.The State Key Laboratory of Refractories and Metallurgy, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, Wuhan, Hubei 430081, P. R. China.The State Key Laboratory of Refractories and Metallurgy, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, Wuhan, Hubei 430081, P. R. China.Research Center of Biomembranomics, State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, P. R. China.The State Key Laboratory of Refractories and Metallurgy, School of Chemistry & Chemical Engineering, Wuhan University of Science and Technology, Wuhan, Hubei 430081, P. R. China.Serine serves as a metabolic nexus in tumors, coordinating one-carbon metabolism, nucleotide synthesis, and redox regulation. While serine transporters (SerTs) are known to be dysregulated in cancer, their functional nanoscale organization remains unresolved due to the limitation of resolution imaging and available probes. Here, we developed a substrate-based fluorescent probe (Ser-probe) enabling direct stochastic optical reconstruction microscopy of SerTs, revealing malignancy-associated clustering assembly of SerTs that correlates with transport capacity. Compared to MDA-MB-231 cells, MCF7 cells with higher endogenous serine biosynthetic capacity exhibited more pronounced SerT/glucose transporter (GluT) co-clustering, suggesting that their spatial assemblies closely correlate with serine transport and biosynthetic functions in maintaining serine homeostasis. Their cluster morphology and co-assembly were revealed to depend critically on lipid rafts and glycan cross-linking, identifying the key determinants of spatial distribution to enable mechanistic understanding and potential regulation. Glucose deprivation weakened SerT/GluT clustering and their colocalization, which may be caused by their attenuated functional cooperativity in serine homeostasis maintenance under glucose-dependent suppression of serine synthesis. Pharmacological inhibition of phosphoglycerate dehydrogenase (PHGDH) initially enhanced SerT/GluT aggregation and colocalization, but this effect gradually attenuated as doses increased. The strategic combination of a PHGDH inhibitor with glucose restriction or free sialic acid synergistically disrupted SerT/GluT nanoscale organization, amplifying the anti-tumor efficacy of the PHGDH inhibitor and establishing the metabolic plasticity of transporter assemblies as a targetable vulnerability. This work establishes a fundamental link between transporter spatial assembly and tumor serine metabolic reprogramming, providing a new perspective to better understand SerT dysfunction in tumor metabolic reprogramming, offering novel therapeutic avenues for targeting serine metabolism in cancer.https://spj.science.org/doi/10.34133/research.0805
spellingShingle Pengwei Jiang
Hao Hou
Jiaqi Wang
Xumin Wang
Yaqi Wang
Simin Liu
Junling Chen
Hongda Wang
Feng Liang
Mechanistic Insight into Serine Flux Regulation through Nanoscale Organization of Glucose and Serine Transporters by Substrate Probe-Based Direct Stochastic Optical Reconstruction Microscopy Imaging
Research
title Mechanistic Insight into Serine Flux Regulation through Nanoscale Organization of Glucose and Serine Transporters by Substrate Probe-Based Direct Stochastic Optical Reconstruction Microscopy Imaging
title_full Mechanistic Insight into Serine Flux Regulation through Nanoscale Organization of Glucose and Serine Transporters by Substrate Probe-Based Direct Stochastic Optical Reconstruction Microscopy Imaging
title_fullStr Mechanistic Insight into Serine Flux Regulation through Nanoscale Organization of Glucose and Serine Transporters by Substrate Probe-Based Direct Stochastic Optical Reconstruction Microscopy Imaging
title_full_unstemmed Mechanistic Insight into Serine Flux Regulation through Nanoscale Organization of Glucose and Serine Transporters by Substrate Probe-Based Direct Stochastic Optical Reconstruction Microscopy Imaging
title_short Mechanistic Insight into Serine Flux Regulation through Nanoscale Organization of Glucose and Serine Transporters by Substrate Probe-Based Direct Stochastic Optical Reconstruction Microscopy Imaging
title_sort mechanistic insight into serine flux regulation through nanoscale organization of glucose and serine transporters by substrate probe based direct stochastic optical reconstruction microscopy imaging
url https://spj.science.org/doi/10.34133/research.0805
work_keys_str_mv AT pengweijiang mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging
AT haohou mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging
AT jiaqiwang mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging
AT xuminwang mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging
AT yaqiwang mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging
AT siminliu mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging
AT junlingchen mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging
AT hongdawang mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging
AT fengliang mechanisticinsightintoserinefluxregulationthroughnanoscaleorganizationofglucoseandserinetransportersbysubstrateprobebaseddirectstochasticopticalreconstructionmicroscopyimaging