| 2021 |
Heterozygous missense variants in RRAGD cause constitutive activation of mTORC1 signaling in vitro, establishing that RagD GTPase mediates amino acid signaling to mTORC1, and that its dysregulation leads to kidney tubulopathy (hypomagnesemia, hypokalemia, nephrocalcinosis) and dilated cardiomyopathy (ADKH-RRAGD syndrome). RagD expression was confirmed along the mammalian nephron including the thick ascending limb and distal convoluted tubule. |
Whole-exome/genome sequencing of patient cohort, in vitro functional analyses of RRAGD variants measuring mTOR signaling activation, immunolocalization along nephron segments |
Journal of the American Society of Nephrology |
High |
34607910
|
| 2023 |
RRAGD auto-activating mutations (causing kidney tubulopathy and cardiomyopathy) constitutively activate a non-canonical mTORC1 signaling branch that specifically phosphorylates TFEB and TFE3 (without affecting canonical substrates like S6K), even in the absence of Folliculin (the GAP responsible for RagC/D activation), thereby inhibiting nuclear translocation and transcriptional activity of TFEB/TFE3 and impairing responses to lysosomal and mitochondrial injury. |
In vitro assays in HeLa and HK-2 cell lines, human iPSC-derived cardiomyocytes, patient-derived primary fibroblasts; TFEB/TFE3 phosphorylation assays; nuclear translocation imaging; Folliculin knockout controls |
Nature Communications |
High |
37188688
|
| 2018 |
TFEB-driven endocytosis promotes assembly of MTORC1-containing nutrient-sensing complexes through formation of endosomes that carry RRAGD, the amino acid transporter SLC38A9, and activate AKT; these TFEB-induced signaling endosomes en route to lysosomes are required to dissociate TSC2 and re-tether/activate MTORC1 on endolysosomal membranes. |
Immunofluorescence colocalization, endocytosis rate assays, MTORC1 activity measurements, TSC2 dissociation assays, overexpression and starvation experiments in CAD and HEK293T cells |
Autophagy |
Medium |
30145926
|
| 2021 |
Loss of FLCN or its binding partners FNIP1/FNIP2 in human renal tubular epithelial cells activates TFE3, which upregulates RRAGD expression (among other E-box targets including GPNMB), without modifying mTORC1 activity, indicating RRAGD is a transcriptional target of TFE3 downstream of FLCN. |
FLCN/FNIP1/FNIP2 knockout in RPTEC/TERT1 cells, RNA-seq, TFE3 ChIP-seq, Western blot, RT-qPCR |
eLife |
High |
33459596
|
| 2023 |
MEF2A and MEF2D transcription factors control the FLCN-FNIP1/2 complex, which acts as a RRAGC-RRAGD GTPase-activating element to promote MTORC1 recruitment to the lysosome and its activation; SRC kinase phosphorylates MEF2D at conserved tyrosine residues to enhance this transcriptional activity and MTORC1 activation. |
ChIP, transcriptional reporter assays, FNIP1/2 knockdown/overexpression, mTORC1 lysosomal localization imaging, MEF2A/D double depletion, SRC phosphorylation site mutagenesis |
Autophagy |
Medium |
37772772
|
| 2024 |
Overexpression of RRAGD patient variants (p.S76L and p.P119R) in zebrafish embryos causes decreased ventricular fractional shortening, reduced ejection fraction, and pericardial swelling; these cardiac phenotypes are reversible with rapamycin (mTOR inhibitor), establishing a direct causal role of RRAGD gain-of-function mutations in cardiomyopathy through mTOR signaling. |
Zebrafish embryo cRNA injection model, echocardiographic measurements (fractional shortening, ejection fraction), rapamycin rescue experiment, survival analysis |
American Journal of Physiology. Heart and Circulatory Physiology |
Medium |
39331021
|
| 2025 |
Novel RRAGD variants p.(Ser77Phe) and p.(Ile100Arg) cause constitutive activation of non-canonical mTORC1 signaling, confirmed by in vitro mTORC1 activity assays; dapagliflozin (SGLT2 inhibitor) treatment in patients with the p.(Thr97Pro) variant increased serum magnesium levels, providing functional evidence linking RRAGD-mTORC1 activity to renal magnesium handling. |
In vitro mTORC1 activity assays for novel variants, clinical evaluation of diuretic and SGLT2 inhibitor treatment in ADKH-RRAGD patients |
Kidney International Reports |
Medium |
41141537
|
| 2025 |
LINC00622 lncRNA associates with and recruits transcription factor BTF3 to the RRAGD promoter, transcriptionally enhancing RRAGD expression, which activates mTORC1 and inhibits autophagic cell death in cutaneous melanoma. |
RNA immunoprecipitation, ChIP, RRAGD promoter reporter assays, LINC00622 knockdown/overexpression with mTORC1 activity readouts and autophagy flux assays |
Cell Death & Disease |
Medium |
40651979
|
| 2022 |
miR-99a-5p directly targets the 3'UTR of RRAGD mRNA (confirmed by dual luciferase reporter assay), negatively regulating RRAGD expression; overexpression of miR-99a-5p inhibits glycolysis (reduced glucose uptake, lactate production, extracellular acidification rate) and induces apoptosis in cervical cancer cells by suppressing RRAGD. |
Dual luciferase reporter assay, RT-qPCR, Western blot, MTT assay, flow cytometry, Seahorse XFe96 extracellular flux analysis, shRNA knockdown |
Oncology Letters |
Medium |
35720506
|
| 2024 |
miR-125b-1-3p directly targets RRAGD, and its overexpression suppresses the RRAGD/mTOR/ULK1 signaling axis, enhancing autophagy in vascular smooth muscle cells, reducing lipid uptake and foam cell formation, and decreasing atherosclerotic plaque development in mice. |
miR-125b-1-3p overexpression in vivo (mice) and in vitro (VSMCs), RRAGD 3'UTR targeting assay (implied interaction), mTOR/ULK1 pathway analysis, autophagy flux assays, atherosclerosis plaque quantification |
Cellular Signalling |
Medium |
38471617
|
| 2021 |
RRAGD knockdown in HCC cell lines reduces glucose uptake, lactate production, and extracellular acidification rate, and inhibits cell proliferation, invasion, and migration; MYC oncogene upregulates RRAGD expression in HCC cells, placing RRAGD downstream of MYC in promoting aerobic glycolysis. |
shRNA knockdown of RRAGD in Huh-7 and HepG2 cells, glucose uptake colorimetric assay, lactate assay, ECAR measurement, Western blot, RT-qPCR for MYC-RRAGD relationship |
Annals of Hepatology |
Medium |
33434687
|
| 2023 |
LncTUG1 acts as a ceRNA sponge for miR-144-3p, relieving miR-144-3p-mediated suppression of RRAGD (miR-144-3p binds 3'UTR of RRAGD mRNA), thereby activating mTOR/S6K pathway and promoting HCC progression; validated in xenograft mouse models showing decreased p-mTOR, p-S6K, and RRAGD upon LncTUG1 knockdown. |
Dual luciferase reporter assay for miR-144-3p/RRAGD 3'UTR interaction, rescue experiments with mTOR pathway inhibitors/activators, xenograft nude mouse models, qRT-PCR, Western blot |
Scientific Reports |
Medium |
37160972
|
| 2025 |
IRTKS forms lysosome-localized membrane-associated condensates that selectively interact with RRAGD (a key upstream regulator of mTORC1), enhancing the sensitivity of mTORC1 to free amino acids; IRTKS-mediated mTORC1 hyperactivation in hepatic knockin mice promotes MASLD and HCC progression. |
Phospho-antibody array screening, co-immunoprecipitation/interaction assay between IRTKS and RRAGD, lysosomal localization imaging, Irtks hepatic knockin mouse model, mTOR activity assays |
Cell Reports |
Medium |
41575860
|
| 2025 |
The CGAS-STING1 pathway activates lysosome biogenesis through lipidated GABARAP on single membranes (regulated by V-ATPase-ATG16L1 axis), which sequesters the FLCN-FNIP complex to abolish its GAP function toward RRAGC and RRAGD, leading to impaired MTORC1-dependent phosphorylation of TFEB and its subsequent nuclear translocation. |
GABARAP lipidation assays, FLCN-FNIP complex sequestration assays, MTORC1 substrate phosphorylation measurements, TFEB nuclear translocation imaging, V-ATPase-ATG16L1 axis dissection |
Autophagy |
Medium |
39835593
|
| 2025 |
The miR-302/367 cluster directly targets RRAGD 3'UTR (validated by dual-luciferase reporter assay), and its overexpression in breast cancer cell lines reduces RRAGD transcript and protein levels, contributing to broad suppression of mTOR pathway activity. |
Dual-luciferase reporter assay, RT-qPCR, Western blot in SK-BR-3 and MDA-MB-231 cells after miR-302/367 vector transfection |
FASEB Journal |
Low |
40817797
|
| 2025 |
RRAGD co-localizes with lysosomal marker LAMP1 and lysosomal regeneration transcription factor TFEB, indicating lysosomal targeting; engineered extracellular vesicles encapsulating RRAGD protein ameliorated lysosomal dysfunction and suppressed apoptosis in nucleus pulposus cells in vitro and in vivo. |
Immunofluorescence colocalization with LAMP1 and TFEB, Fc-TRIM21 engineered EV loading, in vitro and in vivo (intervertebral disc) functional assays |
Journal of Nanobiotechnology |
Low |
41076529
|
| 2025 |
IL4 treatment induces RRAGD expression in follicular lymphoma cells in a STAT6-dependent manner; RRAGD is required for mTOR activation in lymphoma cells, and IL4-enhanced BCR signaling-induced mTOR activation is augmented by mutant STAT6 and reduced by CREBBP mutants through modulation of RRAGD expression. |
RNA-seq gene expression, RRAGD siRNA knockdown with mTOR activity measurement, IL4 stimulation experiments, STAT6 mutant and CREBBP mutant primary FL cells |
Leukemia |
Medium |
39910284
|
| 2024 |
lncARF physically binds to RRAGD protein and inhibits its ubiquitination, thereby activating PI3K/Akt and MAPK signaling pathways downstream of RRAGD; lncARF knockdown decreased atherosclerotic lesion formation by promoting autophagy. |
Mass spectrometry, RNA pull-down, RNA immunoprecipitation (RIP), ubiquitination assays, lncARF knockdown in vivo and in vitro, PI3K/Akt and MAPK pathway activity measurements |
Journal of Advanced Research |
Medium |
39214417
|