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@ARTICLE{Brunialti:270813,
author = {Brunialti, Electra and Rizzi, Nicoletta and Pinto-Costa,
Rita and Villa, Alessandro and Panzeri, Alessia and Meda,
Clara and Rebecchi, Monica and Di Monte, Donato A and Ciana,
Paolo},
title = {{D}esign and validation of a reporter mouse to study the
dynamic regulation of {TFEB} and {TFE}3 activity through in
vivo imaging techniques.},
journal = {Autophagy},
volume = {20},
number = {8},
issn = {1554-8627},
address = {Abingdon, Oxon},
publisher = {Taylor $\&$ Francis},
reportid = {DZNE-2024-00903},
pages = {1879 - 1894},
year = {2024},
note = {ISSN 1554-8635 not unique: **2 hits**.},
abstract = {TFEB and TFE3 belong to the MiT/TFE family of transcription
factors that bind identical DNA responsive elements in the
regulatory regions of target genes. They are involved in
regulating lysosomal biogenesis, function, exocytosis,
autophagy, and lipid catabolism. Precise control of TFEB and
TFE3 activity is crucial for processes such as senescence,
stress response, energy metabolism, and cellular catabolism.
Dysregulation of these factors is implicated in various
diseases, thus researchers have explored pharmacological
approaches to modulate MiT/TFE activity, considering these
transcription factors as potential therapeutic targets.
However, the physiological complexity of their functions and
the lack of suitable in vivo tools have limited the
development of selective MiT/TFE modulating agents. Here, we
have created a reporter-based biosensor, named
CLEARoptimized, facilitating the pharmacological profiling
of TFEB- and TFE3-mediated transcription. This innovative
tool enables the measurement of TFEB and TFE3 activity in
living cells and mice through imaging and biochemical
techniques. CLEARoptimized consists of a promoter with six
coordinated lysosomal expression and regulation motifs
identified through an in-depth bioinformatic analysis of the
promoters of 128 TFEB-target genes. The biosensor drives the
expression of luciferase and tdTomato reporter genes,
allowing the quantification of TFEB and TFE3 activity in
cells and in animals through optical imaging and biochemical
assays. The biosensor's validity was confirmed by modulating
MiT/TFE activity in both cell culture and reporter mice
using physiological and pharmacological stimuli. Overall,
this study introduces an innovative tool for studying
autophagy and lysosomal pathway modulation at various
biological levels, from individual cells to the entire
organism.Abbreviations: CLEAR: coordinated lysosomal
expression and regulation; MAR: matrix attachment regions;
MiT: microphthalmia-associated transcription factor; ROI:
region of interest; TBS: tris-buffered saline; TF:
transcription factor; TFE3: transcription factor binding to
IGHM enhancer 3; TFEB: transcription factor EB; TH: tyrosine
hydroxylase; TK: thymidine kinase; TSS: transcription start
site.},
keywords = {Animals / Basic Helix-Loop-Helix Leucine Zipper
Transcription Factors: metabolism / Mice / Lysosomes:
metabolism / Genes, Reporter / Humans / Promoter Regions,
Genetic: genetics / Autophagy / Biosensing Techniques:
methods / Gene Expression Regulation: drug effects /
Autophagy (Other) / drug discovery (Other) / luciferase
(Other) / lysosomal pathway (Other) / non-invasive study of
TFEB and TFE3 (Other) / optical imaging (Other) / Basic
Helix-Loop-Helix Leucine Zipper Transcription Factors (NLM
Chemicals) / Tcfeb protein, mouse (NLM Chemicals) / Tcfe3
protein, mouse (NLM Chemicals) / TFE3 protein, human (NLM
Chemicals) / TFEB protein, human (NLM Chemicals)},
cin = {AG Di Monte},
ddc = {570},
cid = {I:(DE-2719)1013008},
pnm = {352 - Disease Mechanisms (POF4-352)},
pid = {G:(DE-HGF)POF4-352},
typ = {PUB:(DE-HGF)16},
pmc = {pmc:PMC11262230},
pubmed = {pmid:38522425},
doi = {10.1080/15548627.2024.2334111},
url = {https://pub.dzne.de/record/270813},
}