EDITORIAL www.jasn.

org

mTOR: Pumping Nutrients small GTPases. On the lysosomal membrane, mTORC1 is

directly activated by another small GTPase, Rheb, activity of
into Tubules which is enhanced by the growth factor-PI3K-Akt pathway.
Although aberrant activation of mTORC1 in glomerular
Ken Inoki podocytes leads to their injury/loss and causes glomerular
Life Sciences Institute, Department of Molecular and Integrative dysfunction,5 in renal proximal and distal tubular cells, ab-
Physiology, Division of Nephrology, Department of Internal errant activation of mTORC1 causes their proliferation and
Medicine, University of Michigan, Ann Arbor, Michigan
cystic formation.6 In contrast, loss of functional mTORC1 in
J Am Soc Nephrol 28: cccccc, 2016. podocytes by ablating an essential mTORC1 component,
doi: 10.1681/ASN.2016080924 Raptor, also causes podocyte dysfunction.7 These genetic
observations indicate that both aberrant activation and loss
of mTORC1 activity in podocytes cause vulnerability for
Recent studies have revealed that the mammalian target of
highly differentiated podocytes, which lack a capability of
rapamycin (mTOR) plays an important role in both physio-
proliferation.
logic and pathophysiologic cellular processes in kidney cells.1
Clinical data showed that reduction of serum potassium
mTOR is an evolutionarily conserved protein kinase that forms
and phosphorus and induction of tubular LMW proteinuria
two distinct multiprotein kinase complexes termed mTORC1
can be seen in patients treated with sirolimus (rapamycin)
and mTORC2. Rapamycin is a specic and preponderant al-
after renal transplantation,1,8 suggesting that prolonged
losteric inhibitor for mTORC1. Rapamycin forms a complex
sirolimus treatment causes defects of tubular reabsorption.
with the endogenous FKBP12 protein to interact with the
However, physiologic roles of mTORC1 and/or mTORC2
FKBP12-rapamycin binding (FRB) domain of mTOR kinase
in the regulation of renal tubular absorption system have re-
in mTORC1. Binding this drug-protein complex to the FRB
mained elusive. To investigate roles of each mTORC function
domain blocks the accessibility of substrates to the active site
in renal tubules, Grahammer et al.,9 as described in this issue
of mTOR kinase and ultimately, disrupts the formation of
of the Journal of the American Society of Nephrology (JASN),
mTORC1. A component of mTORC2, Rictor, structurally hin-
generate inducible conditional mTORC1 anord/mTORC2
ders the FRB domain from the rapamycin-FKBP12 complex,
knockout mouse models. Importantly, mice lacking functional
which confers mTORC2 resistant to rapamycin.2 However,
mTORC1 but not mTORC2 in renal tubular cells displayed
prolonged rapamycin treatment often inhibits cellular
glucosuria, phosphaturia, aminoaciduria, low molecular
mTORC2 activity, likely due to a blockade of mTORC2 for-
weight proteinuria, and albuminuria, which are all pathologic
mation through its interaction with newly translated mTOR
phenotypes seen in Fanconi syndrome.
kinase.3 Although a series of potent ATPcompetitive mTOR
Renal Fanconi syndrome is characterized by renal tubular
kinase inhibitors, which inhibit both mTORC1 and mTORC2,
dysfunction and vitamin Dresistant metabolic bone dis-
has recently been developed and is widely used in research,
ease.10 Fanconi syndrome is often due to hereditary autoso-
specic inhibitors for mTORC2 remain underdeveloped.
mal recessive Mendelian disorders, such as cystinosis, type 1
Rapamycin-sensitive mTORC1 plays a key role in stimulating
glycogen storage disease, and Wilson disease. Fanconi syn-
cellular anabolic processes, such as protein and lipid biosynthe-
drome can also be acquired, and it is then most often revers-
sis to support cell growth, differentiation, and proliferation. In
ible and has adult onset. Many metabolic toxins, such as
contrast, it inhibits autophagy, a major catabolic process that
heavy metals, and therapeutic drugs, including tetracycline,
maintains cellular energy and nutrient homeostasis by recycl-
gentamycin, and ifosfamide, cause tubular cell damage and
ing unnecessary and damaged molecules and/or organelles
induce the reabsorptive dysfunctions characteristic of the
through their degradation.
Fanconi syndrome.
mTORC1 activation requires both growth factor and amino
In fact, in mTORC1 knockout mice, uptake of horseradish
acids, especially leucine, arginine, and glutamine.4 Amino
peroxidase and uorescence-labeled lactoglobulin into the
acids, such as leucine, stimulate mTORC1 translocalization
proximal tubules was signicantly reduced compared with
to the lysosomal membrane through the activation of Rag
those in wild-type mice, indicating that the activity of
mTORC1 is required for both uid face and receptor-
mediated endocytosis.9
Published online ahead of print. Publication date available at www.jasn.org.
Proteomics analyses revealed that the expression of cer-
Correspondence: Prof. Ken Inoki, Life Sciences Institutes, 210 Washtenow tain amino acid transporters, such as B0AT1 (Slc6a19), Y1
Avenue, Ann Arbor, MI 48109. Email: inokik@umich.edu
LAT-1 (Slc7a7), and 4F2HC (Slc3a2), was reduced in
Copyright 2016 by the American Society of Nephrology mTORC1 knockout tubules.

J Am Soc Nephrol 28: cccccc, 2016 ISSN : 1046-6673/2801-ccc 1

 EDITORIAL www.jasn.org
In addition, the expression of proteins, such as DOCK8,
RAB10, and sorting nexin 8, proteins important for maintaining
cell polarity and endocytosis, was also mitigated in mTORC1
knockout tubules. However, the expression of major scavenger
receptors, megalin and cubilin, was maintained in mTORC1
knockout tubules.9 Consistent with the in vivo observations,
cultured proximal tubular cells treated with rapamycin or
ATPcompetitive mTOR kinase inhibitors (Torin1 and PP242)
had signicantly reduced endocytosis. Interestingly, PF-
4708671, a specic S6K1 inhibitor, or knockdown of S6K with
the shRNA also blocked endocytosis in proximal tubular cells,
suggesting that loss of S6K1 activity in mTORC1 knockout mice
may cause a reduction of endocytosis in tubular cells. These
observations may also imply that defects of the reabsorption
system in tubular cells in mTORC1 knockout mice are caused by
not only a reduction of functional proteins involved in endocytosis
but also, an inhibition of enzymatic activity of these proteins,
which is likely through their post-translational modications.
Indeed, the study also showed that phosphorylation levels of
certain amino acid transporters (e.g., B0AT1) and a glucose
transporter (SGLT2) were decreased in mTORC1 knockout
tubules. However, it remains unclear whether S6K1 and/or
mTORC1 directly phosphorylate these proteins and stimulate
their functions under physiologic conditions. It will be inter-
esting to determine the activity of endocytosis in the proximal
tubules of S6K1 knockout mice.
Although the data presented in this study clearly indicated
that the physiologic levels of mTORC1 activity are essential for
renal tubular cell functions, the important question from this
study is whether loss of mTORC1 activity is involved in the
mechanisms underlying any inherited or acquired renal Fan-
coni syndrome.
Intriguingly, recent studies have shown that the activity of
mTORC1 is diminished in cystinosis,11,12 which is the most
frequent cause of renal Fanconi syndrome in children. Cysti-
nosis is an autosomal recessive lysosomal storage disorder
caused by mutations in the CTNS gene encoding the lysosomal
cysteine-proton symporter, cystinosin. Loss of functional cys-
tinosin leads to the accumulation of cysteine within the lyso-
some and lysosomal dysfunction, which causes damage in
many tissues and organs, including renal tubular cells. Anti-
gnac and colleagues11 recently showed that cystinosin inter-
acts with vATPases, Ragulator (RagA/B guanine nucleotide
exchange factor), and Rag small GTPases, which are essential
compartments for amino acidinduced mTORC1 recruitment
on the lysosomal membrane for its activation.
Loss of functional cystinosin signicantly blocks amino
acidinduced mTORC1 lysosomal localization and its activation
in mouse proximal tubular cells. Levtchenko and colleagues12
also showed that amino acidinduced mTORC1 activation is
delayed in human proximal tubular cells that lack functional
cystinosin, although they proposed that mTORC1 is rather
retained on the lysosomal membrane under amino acid star-
vation conditions. Although it remains elusive whether forced
mTORC1 activation restores function, including endocytosis,
2 Journal of the American Society of Nephrology
in dysfunctional cystinosin null proximal tubular cells, these
studies and the data described by Grahammer et al.9 in this
issue of the JASN suggest that loss of physiologic mTORC1
activation may be an important mechanism underlying renal
Fanconi syndrome due to lysosomal perturbation.
Additionally, it has been shown that many acquired and
several genetic cases of renal Fanconi syndrome were associated
with mitochondrial dysfunction in renal tubular cells.13,14 The
renal tubular reabsorption system requires robust energy, and
an impairment of ATP production inhibits Na/K ATPase activity,
which is required for maintaining electrochemical sodium gra-
dient across the luminal membrane. Failure of establishing the
sodium gradient decreases the net reabsorption of various solutes
through Na/H exchange, Na/glucose, Na/PO4, and Na/amino acid
cotransporters. Importantly, it has been postulated that intact
mitochondrial function and high concentrations of ATP are
required for the mTOR kinase to phosphorylate its substrates.
Furthermore, the activity of mTORC1 also plays an important
role in mitochondrial biogenesis.15 Thus, the reduction of
intact mitochondrial function may further mitigate the gener-
ation of fresh mitochondria through mTORC1 inhibition.
Consistent with this idea, Grahammer et al.9 observed that
numbers of mitochondria were reduced in mTORC1 knockout
renal tubular cells. Thus, it is conceivable that an impairment in
mitochondrial function seen in certain renal Fanconi syndromes
likely contributes to the reduction of cellular mTORC1 activity in
proximal tubular cells, which may further disturb the capacity of
reabsorption system in the cells. Thus, growing evidence, includ-
ing the results in the work by Grahammer et al.,9 will set the stage
for additional studies to explore the roles of mTORC1 as a sig-
naling nexus in the maintenance of renal proximal tubular cells
and the development of the devastating pathologic consequences
of Fanconi syndrome.
ACKNOWLEDGMENTS
K.I. is supported by grant DK083491 from the National Institute of
Diabetes and Digestive and Kidney Diseases.
DISCLOSURES
None.
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