Anda di halaman 1dari 8

Review

Special Issue: Human Genetics

Worms, bacteria, and micronutrients:


an elegant model of our diet
Lutfu Safak Yilmaz and Albertha J.M. Walhout
Program in Systems Biology, Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA

Micronutrients are required in small proportions in a diet referred to as the microbiota) (Figure 1). The importance
to carry out key metabolic roles for biomass and energy of such indirect effects on human health and disease is
production. Humans receive micronutrients either di- increasingly appreciated [1]. First, microbiota metabolize
rectly from their diet or from gut microbiota that metab- dietary compounds that cannot be metabolized by humans.
olize other nutrients. The nematode Caenorhabditis For instance, gut bacteria break down plant-derived dietary
elegans and its bacterial diet provide a relatively simple fibers resulting in the production of the short-chain fatty
and genetically tractable model to study both direct and acids (SCFAs; see Glossary) propionate and butyrate [2].
microbe-mediated effects of micronutrients. Recently, Second, the microbiota can provide micronutrients such as
this model has been used to gain insight into the rela- vitamin B12 and vitamin K [3]. Third, the breakdown of
tionship between micronutrients, physiology, and me- xenobiotics by microbial metabolism can contribute to the
tabolism. In particular, two B-type vitamins, vitamin B12 nutrient pool in the gut during medical treatment [4]. It is
and folate, have been studied in detail. Here we review therefore perhaps not surprising that the microbiota is
how C. elegans and its bacterial diet provide a powerful increasingly associated with a wide variety of human dis-
interspecies systems biology model that facilitates the eases, including type 2 diabetes [5], cardiovascular disease
precise delineation of micronutrient effects and the [6], obesity [7], immunity and inflammatory disease [8], and
mechanisms involved. even autism [9]. Although the role of the microbiota on
human health and disease is widely acknowledged, the
Diet and micronutrients underlying mechanisms are mostly not resolved.
An organism can be conceptualized as a living system that Systematically studying the effects and mechanisms of
receives dietary inputs and converts these into biomass for individual micronutrients is difficult in humans and other
reproduction, cell renewal, or wound healing, and into en- mammals, including mice. This is not only because of the
ergy to sustain daily tasks at the cellular and organismal long lifespans and internal complexity of these organisms,
levels. The diet consists of macronutrients that are con- but also because genome-scale whole organism genetics is
sumed in large amounts and micronutrients that are much not feasible for ethical reasons or is prohibitively expensive.
less abundant. Macronutrients, such as carbohydrates, fats,
and proteins are the main providers of carbon, nitrogen, and
Glossary
electrons/reducing power and are either incorporated into
biomass or converted to energy. By contrast, the contribu- Life history traits: measurable physiological and phenotypic properties that
affect the health of an animal and the population. Examples include brood size,
tion of micronutrients such as vitamins, cofactors, and most developmental rate, and lifespan.
minerals to biomass and energy generation is more indirect. Postembryonic developmental rate: the rate at which Caenorhabditis elegans
Generally, micronutrients are not metabolized except for proceeds through the larval stages (L1, L2, L3, and L4) to the young adult stage
and then to adulthood.
minor modifications to acquire active molecular forms. How- RNAi: RNA interference to inhibit gene expression. In C. elegans, double-stranded
ever, they do play crucial roles in biomass synthesis RNA (dsRNA) complementary to the targeted mRNA on one strand is given to
worms by feeding a bacterial strain that carries the dsRNA in a plasmid vector.
and energy generation by activating metabolic enzymes,
Short-chain fatty acids (SCFAs): fatty acids with five or fewer carbons,
preventing oxidative damage as antioxidants, or by the including formic- (one carbon), acetic- (two carbons), propionic- (three
hormonal regulation of biological processes. carbons), butyric- (four carbons), and valeric- (five carbons) acid. SCFAs are
products of fatty acid beta oxidation as well as bacterial fermentation, for
Diet can affect the human body directly or indirectly example, in the human colon.
(Figure 1). Direct effects involve the absorbance and utili- Vitamin B12 (cobalamin): one of the water-soluble B vitamins that is important
zation of nutrients from the diet itself. Indirect dietary for blood formation and normal functioning of nerves. It is the most
complicated vitamin, with a cobalt atom placed at the center of a tetra-pyrrole
effects are mediated by the microbial community in the ring. Depending on the group (R) attached to this cobalt atom, it takes different
digestive track, known as the gut microbiota (hereafter forms, which include hydroxocobalamin (R = OH), adenosylcobalamin
(R = adenosyl group), and methylcobalamin (R = methyl group).
Vitamin B9 (folic acid): another water-soluble B vitamin that is important for
Corresponding author: Walhout, A.J.M. (marian.walhout@umassmed.edu). cell division and growth. Folic acid is activated when converted to dihydro-
Keywords: Caenorhabditis elegans; micronutrients; vitamin B; folate; gut microbiota; folate, which is transformed into tetrahydrofolate, the active form in one
metabolism. carbon pool by folate (also known as the folate cycle; see Figure 3 in main text).
0168-9525/ Different derivatives of tetrahydrofolate exist including 10-formyltetrahydrofo-
2014 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.tig.2014.07.010 late, 5,10-methenyltetrahydrofolate, 5,10-methylenetetrahydrofolate, and 5-
methyltetrahydrofolate.

496 Trends in Genetics, November 2014, Vol. 30, No. 11


Downloaded for Tinnaphop Daraswang (tinnaphop8109@gmail.com) at Chulalongkorn University from ClinicalKey.com by Elsevier on May 09, 2017.
For personal use only. No other uses without permission. Copyright 2017. Elsevier Inc. All rights reserved.
Review Trends in Genetics November 2014, Vol. 30, No. 11

Human Worm

Medium

Bacterial diet

Microbiota Key: Macronutrients Drugs


Diet

Intesne Genec
Micronutrients
manipulaon

TRENDS in Genetics

Figure 1. Caenorhabditis elegans as a model organism to study the direct and microbiota-mediated effects of diet.

Further, gut microbiota are highly complex communities to directly absorb micronutrients and drugs from its
including many species that are yet to be cultured in the environment or the growth medium, representing direct
laboratory. Here, we discuss how the nematode C. elegans effects by itself.
and its bacterial diet have been used as a genetically trac- The standard laboratory diet of C. elegans has been OP50,
table model to gain insights into the physiological effects of a B strain of Escherichia coli, although it has been fed many
micronutrients, with a focus on B-type vitamins. different types of bacteria in controlled experiments [16].
Typically, a co-culture of C. elegans and a bacterium is grown
C. elegans as a model organism on solid media supplemented with peptone to promote
C. elegans is a metazoan that lives in temperate soil bacterial proliferation, and this mode of growth is referred
environments and mainly feeds on bacteria growing on to as monoxenic. However, C. elegans can also be cultured in
rotting vegetation [10]. Since it was introduced as a genetic liquid with bacteria, or in an axenic medium with a well-
model organism [11], the worm has proven to be a power- defined set of macro- and micronutrients [17]. Thus, the
ful model that has served to provide seminal insights into a direct effects of specific nutrients on C. elegans can be tested
myriad of biological processes including development by the addition or omission of these factors either in axenic
[12,13], cancer [14], and aging [15]. C. elegans offers mul- or monoxenic growth conditions. By contrast, postembryonic
tiple unique advantages as a model organism, such as developmental rate and fecundity are compromised in axe-
physiological simplicity (1000 somatic cells), a short life- nic medium and in monoxenic conditions with dead bacteria
span (2 weeks), an early and short reproductive period, [18,19]. This implies that metabolically active bacteria sup-
and hermaphroditic reproduction. port optimal worm health, analogous to the microbiota that
In recent years, the worm has also been developed as a grows in the nutritionally rich environment of the human
suitable model to study the effects of diet on metabolism, gut and supports human health by metabolic byproducts.
physiology, and gene expression. An interesting aspect of Because C. elegans and its bacterial diets are genetically
the C. elegans model is that bacteria serve as food, which tractable, they can both be manipulated to systematically
means that bacterial biomass is the main source of macro- dissect the underlying mechanisms using an interspecies
nutrients. Thus, the relationship of C. elegans with bacte- systems biology approach.
ria is different from the synergistic relationship between Different bacterial diets cause significant changes in
mammals and microbiota. However, live bacteria can also C. elegans physiology and gene expression when compared
provide sufficient supplies of micronutrients to the worm to the standard OP50 diet [1922]. Changes elicited by one
as a byproduct of their metabolism, absorb compounds particular bacterial diet, Comamonas aquatica (hereafter
from the environment and deliver them to the worm, or referred to as Comamonas), were attributed to a dilutable
convert these compounds into other molecules that are compound, because this species can affect worm develop-
useful for the worm. Thus, the bacterial component of the ment and gene expression even when dramatically diluted
C. elegans model can represent both direct and indirect with the OP50 diet [19]. Combined genetic analyses of
aspects of a diet (Figure 1). In addition, C. elegans is able E. coli, Comamonas, and the worm itself unveiled that
497
Downloaded for Tinnaphop Daraswang (tinnaphop8109@gmail.com) at Chulalongkorn University from ClinicalKey.com by Elsevier on May 09, 2017.
For personal use only. No other uses without permission. Copyright 2017. Elsevier Inc. All rights reserved.
Review Trends in Genetics November 2014, Vol. 30, No. 11

this dilutable compound is the micronutrient vitamin B12


[23] (see below). This provides a powerful example of how Propionic acid
monoxenic growth of C. elegans can be used as a model Odd-
chain fay acids Amino acids
system to derive connections between micronutrients, gene
expression, and life history traits. We will first review
Mitochondria Cytosol
studies of vitamin B12 and folate using the worm model,
and then briefly discuss additional studies that focused on ppcoa
other micronutrients. pcca-1
pccb-1
Vitamin B12 profoundly affects C. elegans physiology D-mmcoa
and metabolism
mce-1 R-Cbl
Vitamin B12, or cobalamin, occurs in different forms and is (Vitamin B12)
an essential cofactor for two enzymes in two distinct path- L-mmcoa
cblc-1
ways. The first enzyme is methylmalonyl coenzyme A Y76A2B.5 R
mmcm-1
(CoA) mutase (MCM), which uses adenosylcobalamin as MCM adoCbl
mmab-1
Cbl (II)
a cofactor and converts methylmalonyl-CoA to succinyl-
CoA as part of the mitochondrial propionic acid breakdown succoa
pathway (Figure 2). This pathway receives carbon from the
catabolism of propionic acid, branched chain amino acids,
odd-chain fatty acids, and cholesterol. The end product TCA cycle
succinyl-CoA then enters the tricarboxylic acid (TCA) cycle
to generate energy and recycle carbon. The interruption of
this pathway in humans, either due to recessive mutations Energy Biosynthesis
in the gene encoding MCM or because of vitamin B12 Glycolysis
deficiency, causes the devastating disease methylmalonic
aciduria, which manifests itself as organ failure as a result TRENDS in Genetics

of the accumulation of methylmalonic acid in tissues and Figure 2. The propionic acid breakdown pathway and vitamin B12. Caenorhabditis
body fluids [2426]. elegans genes that are associated with each reaction are shown in italics. Human
orthologs of selected genes (bold italics) are provided in Table 1. Broken arrows
The second vitamin B12-dependent enzyme, methio-
indicate pathways not shown in detail. Abbreviations: MCM, methylmalonyl-CoA
nine synthase (MS), uses methylcobalamin to convert mutase; ppcoa, propionyl CoA; mmcoa, methylmalonyl CoA; succoa, succinyl
homocysteine (HCys) to methionine in the cytosolic methi- CoA; Cbl, cobalamin; adoCbl, adonesylcobalamine.
onine/S-adenosylmethionine (SAM) cycle (Figure 3). Be-
cause of this reaction, vitamin B12 deficiency is associated

Folt-1
RFC

R-Cbl (Vitamin B12) Folate


cblc-1
R dhfr-1
Cbl (II) THF DHF
Y76A2B.5 Ser

SAH ahcy-1 HCys dTMP


meCbI Purine mel-32
R-CH3 synthesis Gly
Y110A7A.4
metr-1
mtrr-1

MS F38B6.4
K07E.34 dUMP
C55F2.1
dao-3
R
Cbl (I) mlTHF
SAM sams-1 Met 10fTHF meTHF
sams-3
mthf-1
sams-4
sams-5
5mTHF
TRENDS in Genetics

Figure 3. Methionine and folic acid cycles. Caenorhabditis elegans genes that are associated with each reaction are shown in italics. Human orthologs of selected genes
(bold italics) are provided in Table 1. Broken arrows indicate multiple reactions with multiple genes. Abbreviations: MS, methionine synthase; RFC, reduced folate carrier;
DHF, dihydrofolate; THF, tetrahydrofolate; 10fTHF, 10-formyltetrahydrofolate; meTHF, 5,10-methenyltetrahydrofolate; mlTHF, 5,10-methylenetetrahydrofolate; 5mTHF, 5-
methyltetrahydrofolate; HCys, homocysteine; Met, methionine; SAM, S-adenosyl methionine; SAH, S-adenosyl homocysteine; Cbl, cobalamin; meCbl, methylcobalamine;
Ser, serine; Gly, glycine.

498
Downloaded for Tinnaphop Daraswang (tinnaphop8109@gmail.com) at Chulalongkorn University from ClinicalKey.com by Elsevier on May 09, 2017.
For personal use only. No other uses without permission. Copyright 2017. Elsevier Inc. All rights reserved.
Review Trends in Genetics November 2014, Vol. 30, No. 11

with the metabolic syndrome homocysteinemia, which Watson et al. [23] recently discovered that the standard
results from the toxic build-up of HCys [27,28]. MS activity E. coli diet does not represent ample vitamin B12 conditions,
is connected to DNA synthesis and amino acid interconver- although animals may not be truly vitamin B12 deficient
sions via the folic acid cycle (Figure 3). Some of the methi- [35]. They found that the acceleration of worm development
onine synthesized by MS is used for protein production, and reduction in brood size elicited by a Comamonas diet
while the rest is recycled back to HCys by the generation could be partly explained by this bacteriums ability to
and subsequent conversion of SAM into S-adenosylhomo- provide higher levels of vitamin B12. Previously, they had
cysteine (SAH). SAM acts as the major methyl donor for the generated a C. elegans dietary sensor strain that expresses
methylation of nucleic acids, proteins (e.g., histones), lipids the green fluorescent protein (GFP) under the control of the
(e.g., phosphatidylcholine), and other metabolites such as promoter of the most differentially expressed gene, the acyl-
neurotransmitters, creatine, carnitine, and polyamines. CoA dehydrogenase acdh-1, which serves as a proxy for the
Thus, MS activity can be linked to important cellular induced gene expression changes [19]. On animals fed E.
processes involving biomass synthesis as well as regula- coli, GFP expression is high, whereas on Comamonas it is
tion. In accordance with these broad impacts of MS barely detectable. By transposon-based mutagenesis in
(Figure 3), vitamin B12 deficiency has also been associated Comamonas, Watson et al. identified mutations in the vita-
with megaloblastic anemia [29], neuropathy [30], fetal min B12 biosynthesis pathway that lead to a failure of C.
development defects [31], and pregnancy loss [32]. Overall, elegans to repress GFP expression [23]. In addition, a ge-
vitamin B12 is a crucial micronutrient for human health nome-scale screen of the E. coli deletion collection [43]
because of its role in both pathways (Figures 2 and 3). showed that a deletion in the tonB gene further activates
Whereas many insects and plants do not need vitamin GFP expression in the worm. This gene encodes an out-
B12, C. elegans, like humans, must obtain vitamin B12 ermembrane siderophore transporter that is important for
from its diet. Remarkably, vitamin B12 is uniquely syn- the uptake of vitamin B12 by E. coli. Furthermore, RNAi
thesized by bacteria and archaea [33], with the better silencing of two worm genes involved in the conversion of
defined bacterial pathway comprising more than 20 cobalamin to its active forms, mttr-1 and Y76A2B.5
enzymes. It is estimated that about a third of bacterial (Figure 2, Table 1), repressed the response of the dietary
species can synthesize vitamin B12 [34]. Because E. coli sensor. Also, genes encoding the enzymes that directly use
does not have this pathway, the source of vitamin B12 in vitamin B12 as a cofactor, metr-1 and mmcm-1, were both
the standard C. elegans diet is the vitamin absorbed by uncovered during prior genome-scale genetic screens in the
bacteria from culturing media. worm using the dietary sensor [44]. Consistent with all the
The requirement for exogenously provided vitamin B12 genetic evidence, direct supplementation of the standard E.
for C. elegans was recently demonstrated. Bito et al. prop- coli diet with extra vitamin B12 both repressed the dietary
agated worms for several generations on peptone-free sensor and, importantly, mimicked the effects of the Coma-
medium with bacteria grown in the absence of vitamin monas diet on developmental rate and egg laying. However,
B12 to establish truly vitamin B12-deficient conditions the negative effect of Comamonas on C. elegans lifespan [19]
[35]. They then measured various physiological variables was not reproduced by vitamin B12, demonstrating that
in the worm, as well as enzymatic activity of MS and MCM, other bacterial factors are responsible for this effect.
and the concentration of key metabolites. In agreement To determine the mechanism of developmental acceler-
with human and mouse studies showing fetal loss ation, Watson et al. tested the effect of vitamin B12 on
and infertility under vitamin B12 deplete conditions
[32,36,37], the brood size of vitamin B12-deficient worms
Table 1. Selected genes in Vitamin B12- and folic acid-related
was significantly reduced. In addition, the average lifespan pathwaysa
was reduced and animals were shorter. Not surprisingly,
Pathway Protein name Homo Caenorhabditis
these observations were correlated with loss of activity in figure sapiens elegans
MS and MCM and a corresponding increase in methylma- Figure 2 Propionyl-CoA PCCA pcca-1
lonic acid and HCys levels. Because the latter can inhibit carboxylase alpha
collagen biosynthesis [38], the authors hypothesized that subunit
the adverse effects of vitamin B12 deficiency on C. elegans Propionyl-CoA PCCB pccb-1
carboxylase beta
morphology were due to increased HCys levels. Further-
subunit
more, the authors indicated that the SAM/SAH ratio is Methylmalonyl-CoA MCEE mce-1
altered during vitamin B12 deficiency in the worm epimerase
(Figure 3). Although DNA methylation does not occur in Methylmalonyl-CoA MCM mmcm-1
worms, histone methylation does [39], and therefore vita- mutase
min B12 deficiency can potentially affect gene expression Figure 3 Methionine synthase MTRR mtrr-1
via this pathway. Moreover, because SAM is a highly reductase
anabolic molecule, a reduction in vitamin B12 can also Methionine synthase MTR metr-1
S-Adenosylmethionine MAT1A sams-1
affect biomass production directly by decreasing the syn-
synthetase
thesis of essential components such as phosphatidylcholine Reduced folate carrier SLC19A1 folt-1
in cellular membranes [40]. Probably because of the regu- Figures 2 Methylmalonic aciduria MMADHC Y76A2B.5
latory and metabolic roles of SAM, sams genes in C. elegans and 3 and homocystinuria
(Figure 3) have been associated with fertility [41] and type D protein
aging [42]. a
See Figures 2 and 3 for the function of genes in these pathways.

499
Downloaded for Tinnaphop Daraswang (tinnaphop8109@gmail.com) at Chulalongkorn University from ClinicalKey.com by Elsevier on May 09, 2017.
For personal use only. No other uses without permission. Copyright 2017. Elsevier Inc. All rights reserved.
Review Trends in Genetics November 2014, Vol. 30, No. 11

animals that harbor mutations in either of the two path- Similar to humans, C. elegans obtains folate from its
ways that require this vitamin (Figures 2 and 3). Although diet. Earlier studies with the worm focused on folate
the developmental rate of animals harboring mutations in transport: the folt-1 gene was found to encode the major
mce-1 or pcca-1 (propionic acid breakdown; Table 1, folate uptake system [52]. This resembles the correspond-
Figure 2) could be accelerated by vitamin B12 supplemen- ing protein in mammals, the reduced folate carrier (RFC;
tation, mutations in sams-1 or metr-1 (methionine/SAM Figure 3, Table 1), in multiple ways. First, both proteins
cycle; Table 1, Figure 3) rendered the animal unresponsive are able to transport not only folate but also its derivatives.
to vitamin B12. The effects of these mutations on egg Secondly, FOLT-1 and mammalian RFC are both develop-
laying were similar. Thus, the vitamin B12 phenotypes mentally regulated. The expression of folt-1 is highest in
related to biomass production occur via the methionine/ earlier larval stages [52]. Similarly, in rats, RFC is
SAM cycle, which is in agreement with the central anabolic expressed at higher levels during earlier stages of post-
role of this pathway (Figure 3). embryonic development [53]. Finally, drugs that inhibit
A major function of the propionic acid breakdown folate uptake by targeting RFC in humans also do so in the
pathway (Figure 2) is to prevent build-up of this toxic worm [52].
metabolite, which can be produced by central metabolism Because studies with RFC in mammals had been lim-
or obtained from the diet. In propionic acid toxicity assays ited to the cellular level, Balamurugan et al. [52] proposed
[23], vitamin B12 supplementation significantly im- that C. elegans may provide a suitable model to study
proved the animals ability to withstand high doses of unknown aspects of folate uptake at the whole animal
propionic acid, presumably by enhancing the catabolism level. Austin et al. [54] found that loss of folt-1 reduces C.
of propionyl-CoA (Figure 2). Interestingly, a Dmetr-1 elegans lifespan by 44%, and greatly reduces fertility by
mutant (Table 1, Figure 3) had improved survival rates attenuating spermatogenesis and decreasing oogenesis
under vitamin B12-limiting conditions as compared to probably as a result of the reduction in germline nuclei
wild type animals, implicating that the two vitamin B12 [54]. In addition, the defecation rate of Dfolt-1 animals is
reactions may compete with each other for this cofactor, lower than that of wild type animals, which suggests an
and that blockage of one pathway may increase the flux in overall reduced metabolic rate. Thus folate uptake seems
the other. In humans, propionic acid is produced by the to play an important role in both reproduction and me-
microbiota from the digestion of dietary fibers [2] as well tabolism of C. elegans. However, because the deletion
as by endogenous metabolism from propionyl-CoA. Al- allele used in this study was not outcrossed, in the ab-
though propionic acid has positive roles in human metab- sence of complementation tests to rescue the mutation
olism such as serving as an energy source [2], high levels and additional RNAi tests, there is also the possibility
of propionate have been associated with toxicity, leading that unknown linked mutations explain the observed
to neurological disorders and gingival inflammation [45]. phenotypes.
The inborn metabolic disorder propionic aciduria/acide- Other recent studies on C. elegans folate metabolism
mia is an extreme case of toxicity where mutations in the emphasized the indirect effects of diet (Figure 1). For
PCCA gene (Table 1) render the enzyme inactive thereby instance, Virk et al. [55] serendipitously discovered that
causing the build-up of propionate and other toxic com- impairment of folate synthesis in E. coli extends C. elegans
pounds resulting in damage to various organs and even- lifespan by more than 30%. This can occur genetically by
tually death [46]. The worm results indicate that this mutating a key bacterial gene (aroD) that is involved in the
animal is also a good model to study propionic acid biosynthesis of aromatic compounds such as precursors of
toxicity [23]. folate, or by inhibiting folate synthesis with the bacterio-
static antibiotic sulfamethoxazole. Supplementation with
Vitamin B9, or folate, greatly affects C. elegans biology folate precursors overrides the effect of aroD mutants and
by direct or microbe-mediated mechanisms restores lifespan. Analytical measurements showed that
Folic acid (or folate) is another water-soluble B vitamin. the folate cycle in the worm was functional even when
The active forms of folate act as donors or acceptors of one- bacterial biosynthesis of folate is blocked. Although reduc-
carbon groups in amino acid and nucleic acid metabolism tion of bacterial folate synthesis delays aging, lifespan is
(Figure 3). Owing to its role in the biosynthesis of thymine reduced in folt-1 mutant animals [54] (see above). Thus,
and purines, as well as in amino acid conversions, folate there must be an optimal level of folate in the worm to
levels are critical for fast-dividing cells. Accordingly, folate support long lifespan because both limited and excess
deficiency in humans has been associated with anemia folate can reduce lifespan.
[47], fetal growth retardation [48], and neural tube defects Although bacterial growth was not affected by drug
during embryonic development [49]. Folate and vitamin treatment under the conditions tested [55], it is still not
B12 metabolism intersect in the methionine/SAM cycle clear if the observed changes are due to broader impacts of
(Figure 3); it is probably because of this intersection that folate metabolism in the bacterium, or simply to a decrease
these two vitamins can have similar effects [50]. For in the amount of dietary folate [56]. Experiments with
instance, vitamin B12 deficiency can mimic folate deficien- axenic medium where bacterial metabolism is eliminated
cy because of the so-called methyl trap [51], where 5- will be helpful to directly test the effect of dietary folate on
methyltetrahydrofolate (5mTHF) cannot be converted the worm. An interesting observation in support of bacte-
back to tetrahydrofolate (THF) as a result of the lowered rial folate synthesis being the main effector is that
activity of MS, and therefore, active folate forms cannot be sulfo drugs that inhibit microbial folate biosynthesis also
replenished. extend lifespan in rats [57]. Taken together, C. elegans is a
500
Downloaded for Tinnaphop Daraswang (tinnaphop8109@gmail.com) at Chulalongkorn University from ClinicalKey.com by Elsevier on May 09, 2017.
For personal use only. No other uses without permission. Copyright 2017. Elsevier Inc. All rights reserved.
Review Trends in Genetics November 2014, Vol. 30, No. 11

suitable model to derive hypotheses for the effects of folate the production of fermentation metabolites such as etha-
metabolism status in microbiota on human health [55,56]. nol and lactic acid that may be the actual causes of lifespan
A surprising microbe-mediated effect on worm physi- extension in the worm [73]. However, more recently,
ology and metabolism was recently discovered [58] dur- Gomez et al. [74] showed that lactic acid produced by Q-
ing the studies with the anti-diabetic drug metformin, less E. coli was not responsible for the life-span extension.
which had been known to extend lifespan in C. elegans Instead, they suggested that the survival rate of worms
[59] and mice [60]. Cabreiro et al. showed that this occurs may be improved due to the delay in the colonization of the
through bacterial folate and methionine metabolism [58]. worm gut by respiratory-deficient bacteria as compared to
Remarkably, metformin increases the lifespan of C. ele- the wild type bacteria.
gans when fed live bacteria, but reduces it when animals Finally, a surprising compound that was recently shown
are fed dead bacteria or are grown in axenic medium. to delay aging in C. elegans is alpha-ketoglutarate (AKG)
Analysis of active bacteria showed that metformin chan- [75], an intermediate in the TCA cycle. Direct supplemen-
ged the relative abundance of folate derivatives: meTHF tation of AKG extended lifespan by more than 25% irre-
was increased, whereas methionine was decreased, spective of bacterial metabolic activity. Rather, AKG acts
which is consistent with a possible inhibition of MS by binding and suppressing ATP synthase to reduce energy
(Figure 3). The authors proposed that decreased methio- generation in the worm. This effect is dependent on the
nine levels in the diet reduced the SAM/SAH ratio in the target of rapamycin pathway, whose inactivation delays
worm (as can be inferred from Figure 3), which mimics aging [76]. Overall, AKG supplementation creates DR-like
dietary restriction (DR). Glucose supplementation sup- conditions. Interestingly, AKG concentrations also in-
pressed the effects of metformin in C. elegans, suggesting crease during starvation, suggesting the presence of a
that a high sugar diet might offset the benefits of met- feedback mechanism responding to nutrient availability
formin [58]. Taken together, bacteria may mediate some [75]. Thus the common metabolite AKG also acts as a
of the effects of metformin, which has implications for signaling molecule.
human treatment as well because it may affect or be
metabolized by the microbiota. This is supported by the Concluding remarks
observation that the composition of microbiota is sensi- Taken together, the studies we discussed firmly establish
tive to metformin treatment in rats [61,62] and type 2 the nematode C. elegans and its bacterial diet as a model
diabetes patients [63]. for both direct and microbe-mediated effects of diet on
health and longevity. Because micronutrients obtained
Other micronutrients from the diet or supplied by the microbiota affect human
Micronutrients other than B vitamins have also been health by complicated mechanisms that are difficult to
studied for their direct or bacteria-mediated effects on C. systematically characterize, it is likely that studies using
elegans, particularly on aging. Based on the mitochondrial the interspecies worm system will greatly illuminate basic
theory of aging [64], antioxidants such as vitamin C and physiological responses and underlying mechanisms.
vitamin E may be expected to increase lifespan, but studies The worm/bacteria system will not be a suitable model
with these compounds disagree on their effects on humans for all aspects of human diet and metabolic diseases. First,
as well as C. elegans [65,66]. Because the bacterial diet of C. elegans and the co-cultured bacteria do not represent a
C. elegans is not a source of these vitamins, they are to be case of mutualism as in human and microbiota, but have a
tested by dietary supplementation. However, worms can- trophic relationship. Thus, the C. elegans/bacteria model is
not efficiently uptake these compounds [65,66]. Recently, limited to a simplified conceptual model of direct and
Nishikawa and colleagues developed methods to orally indirect effects as in Figure 1. Second, whereas human
deliver to C. elegans both hydrophilic compounds such microbiota is a complex community, C. elegans interacts
as vitamin C derivatives [67] and hydrophobic compounds with a pure culture during monoxenic growth in the labo-
such as vitamin E derivatives [68] by vehicles that act as ratory. However, monoxenically grown C. elegans does
bacteria-like particles. For both families of vitamins, they offer the possibility of testing different bacterial species
were able to demonstrate an increase in lifespan, thus individually for their role on indirect dietary effects, as
demonstrating the direct effect of these antioxidants for long as they can be cultured in the laboratory. In any case,
the worm model. pathway-level mechanisms discovered in the worm can
Another micronutrient that may affect aging in serve as hypotheses for further testing in mammalian
C. elegans is coenzyme Q (Q), which has diverse functions model systems or humans.
including its major role in the electron transport chain and
as an antioxidant [69]. C. elegans produces its own specific Acknowledgments
form (Q9) [70], but can also use Q8 from its E. coli diet [71]. This work was supported by the National Institutes of Health (grant
When fed mutant E. coli that do not produce Q8, the DK068429 to A.J.M.W.). We thank members of the Walhout laboratory,
lifespan of the worm increases [72]. Remarkably, this notably Lesley MacNeil and Emma Watson, for helpful discussions. We
are also grateful to the anonymous reviewers for their valuable input.
effect was the result of altered bacterial metabolism,
and not due to direct dietary contribution [73], because
supplemented Q10 did not reverse the lifespan extension. References
1 Sommer, F. and Backhed, F. (2013) The gut microbiota masters of
Further, the lifespan extension with Q-less E. coli was not host development and physiology. Nat. Rev. Microbiol. 11, 227238
due to lowered caloric intake because of compromised 2 Tremaroli, V. and Backhed, F. (2012) Functional interactions between
bacterial growth. One possibility for the mechanism is the gut microbiota and host metabolism. Nature 489, 242249

501
Downloaded for Tinnaphop Daraswang (tinnaphop8109@gmail.com) at Chulalongkorn University from ClinicalKey.com by Elsevier on May 09, 2017.
For personal use only. No other uses without permission. Copyright 2017. Elsevier Inc. All rights reserved.
Review Trends in Genetics November 2014, Vol. 30, No. 11

3 LeBlanc, J.G. et al. (2013) Bacteria as vitamin suppliers to their host: a 32 Candito, M. et al. (2003) Clinical B12 deficiency in one case of recurrent
gut microbiota perspective. Curr. Opin. Biotech. 24, 160168 spontaneous pregnancy loss. Clin. Chem. Lab. Med. 41, 10261027
4 Ursell, L.K. et al. (2014) The intestinal metabolome: an intersection 33 Rodionov, D.A. et al. (2003) Comparative genomics of the vitamin B12
between microbiota and host. Gastroenterology 146, 14701476 metabolism and regulation in prokaryotes. J. Biol. Chem. 278, 41148
5 Tilg, H. and Moschen, A.R. (2014) Microbiota and diabetes: an evolving 41159
relationship. Gut 63, 15131521 34 Sanudo-Wilhelmy, S.A. et al. (2014) The role of B vitamins in marine
6 Wang, Z.N. et al. (2011) Gut flora metabolism of phosphatidylcholine biogeochemistry. Annu. Rev. Mar. Sci. 6, 339367
promotes cardiovascular disease. Nature 472, 5763 35 Bito, T. et al. (2013) Vitamin B12 deficiency in Caenorhabditis elegans
7 Million, M. et al. (2013) Gut bacterial microbiota and obesity. Clin. results in loss of fertility, extended life cycle, and reduced lifespan.
Microbiol. Infect. 19, 305313 FEBS Open Bio. 3, 112117
8 Kamada, N. et al. (2013) Role of the gut microbiota in immunity and 36 Pront, R. et al. (2009) Prevalence of low serum cobalamin in infertile
inflammatory disease. Nat. Rev. Immunol. 13, 321335 couples. Andrologia 41, 4650
9 Hsiao, E.Y. et al. (2013) Microbiota modulate behavioral and 37 Watanabe, T. et al. (2003) The effects of dietary vitamin B12 deficiency
physiological abnormalities associated with neurodevelopmental on sperm maturation in developing and growing male rats. Congenit.
disorders. Cell 155, 14511463 Anom. 43, 5764
10 Felix, M.A. and Braendle, C. (2010) The natural history of 38 Thaler, R. et al. (2011) Homocysteine suppresses the expression of the
Caenorhabditis elegans. Curr. Biol. 20, R965R969 collagen cross-linker lysyl oxidase involving IL-6, Fli1, and epigenetic
11 Brenner, S. (1974) The genetics of Caenorhabditis elegans. Genetics 77, DNA methylation. J. Biol. Chem. 286, 55785588
7194 39 Greer, E.L. et al. (2014) A histone methylation network regulates
12 Emmons, S.W. (2005) Male development. In WormBook (Meyer, B.J., transgenerational epigenetic memory in C. elegans. Cell Rep. 7,
ed.), The C. elegans Research Community (http://www.wormbook.org) 113126
13 Sternberg, P.W. (2005) Vulval development. In WormBook (Meyer, 40 Walker, A.K. et al. (2011) A conserved SREBP-1/phosphatidylcholine
B.J., ed.), The C. elegans Research Community (http://www. feedback circuit regulates lipogenesis in metazoans. Cell 147, 840852
wormbook.org) 41 Tamiya, H. et al. (2013) Conserved SAMS function in regulating egg-
14 Kirienko, N.V. et al. (2010) Cancer models in Caenorhabditis elegans. laying in C. elegans. J. Recept. Signal Transduct. Res. 33, 5662
Dev. Dyn. 239, 14131448 42 Hansen, M. et al. (2005) New genes tied to endocrine, metabolic, and
15 Kenyon, C.J. (2010) The genetics of ageing. Nature 464, 504512 dietary regulation of lifespan from a Caenorhabditis elegans genomic
16 Avery, L. and You, Y.J. (2012) C. elegans feeding. In WormBook RNAi screen. PLoS Genet. 1, 119128
(Jorgensen, E.M., ed.), The C. elegans Research Community (http:// 43 Baba, T. et al. (2006) Construction of Escherichia coli K-12 in-frame,
www.wormbook.org) single-gene knockout mutants: the Keio collection. Mol. Syst. Biol. 2,
17 Lu, N.C. and Goetsch, K.M. (1993) Carbohydrate requirement of 2006.0008
Caenorhabditis elegans and the final development of a chemically- 44 Watson, E. et al. (2013) Integration of metabolic and gene regulatory
defined medium. Nematologica 39, 303311 networks modulates the C. elegans dietary response. Cell 153, 253266
18 Lenaerts, I. et al. (2008) Dietary restriction of Caenorhabditis elegans 45 Al-Lahham, S.H. et al. (2010) Biological effects of propionic acid in
by axenic culture reflects nutritional requirement for constituents humans; metabolism, potential applications and underlying
provided by metabolically active microbes. J. Gerontol. A: Biol. Sci. mechanisms. Biochim. Biophys. Acta 1801, 11751183
Med. Sci. 63, 242252 46 Pena, L. et al. (2012) Natural history of propionic acidemia. Mol. Genet.
19 MacNeil, L.T. et al. (2013) Diet-induced developmental acceleration Metab. 105, 59
independent of TOR and insulin in C. elegans. Cell 153, 240252 47 Odewole, O.A. et al. (2013) Near-elimination of folate-deficiency
20 Coolon, J.D. et al. (2009) Caenorhabditis elegans genomic response to anemia by mandatory folic acid fortification in older US adults:
soil bacteria predicts environment-specific genetic effects on life history Reasons for Geographic and Racial Differences in Stroke study
traits. PLoS Genet. 5, e1000503 2003-2007. Am. J. Clin. Nutr. 98, 10421047
21 Shtonda, B.B. and Avery, L. (2006) Dietary choice behavior in 48 van Uitert, E.M. and Steegers-Theunissen, R.P. (2013) Influence of
Caenorhabditis elegans. J. Exp. Biol. 209, 89102 maternal folate status on human fetal growth parameters. Mol. Nutr.
22 Soukas, A.A. et al. (2009) Rictor/TORC2 regulates fat metabolism, Food Res. 57, 582595
feeding, growth, and life span in Caenorhabditis elegans. Genes Dev. 49 Osterhues, A. et al. (2013) The role of folic acid fortification in neural
23, 496511 tube defects: a review. Crit. Rev. Food Sci. Nutr. 53, 11801190
23 Watson, E. et al. (2014) Interspecies systems biology uncovers 50 Shane, B. and Stokstad, E.L. (1985) Vitamin B12-folate
metabolites affecting C. elegans gene expression and life history interrelationships. Annu. Rev. Nutr. 5, 115141
traits. Cell 156, 759770 51 Mato, J.M. et al. (2008) Methionine metabolism and liver disease.
24 Hauser, N.S. et al. (2011) Variable dietary management of Annu. Rev. Nutr. 28, 273293
methylmalonic acidemia: metabolic and energetic correlations. Am. 52 Balamurugan, K. et al. (2007) Cloning and functional characterization
J. Clin. Nutr. 93, 4756 of a folate transporter from the nematode Caenorhabditis elegans. Am.
25 Horster, F. et al. (2009) Prediction of outcome in isolated J. Physiol. Cell Physiol. 293, C670C681
methylmalonic acidurias: combined use of clinical and biochemical 53 Balamurugan, K. and Said, H.M. (2003) Ontogenic regulation of folate
parameters. J. Inherit. Metab. Dis. 32, 630639 transport across rat jejunal brush-border membrane. Am. J. Physiol.
26 Weisfeld-Adams, J.D. et al. (2010) Newborn screening and early Gastrointest. Liver Physiol. 285, G1068G1073
biochemical follow-up in combined methylmalonic aciduria and 54 Austin, M.U. et al. (2010) Knockout of the folate transporter folt-1
homocystinuria, cblC type, and utility of methionine as a secondary causes germline and somatic defects in C. elegans. BMC Dev. Biol. 10,
screening analyte. Mol. Genet. Metab. 99, 116123 http://dx.doi.org/10.1186/1471-213X-10-46
27 Deshmukh, U.S. et al. (2010) Effect of physiological doses of oral 55 Virk, B. et al. (2012) Excessive folate synthesis limits lifespan in the C.
vitamin B-12 on plasma homocysteine: a randomized, placebo- elegans: E. coli aging model. BMC Biol. 10, http://dx.doi.org/10.1186/
controlled, double-blind trial in India. Eur. J. Clin. Nutr. 64, 495502 1741-7007-10-67
28 Remacha, A.F. et al. (2011) Vitamin B12 deficiency, 56 Nguyen, T.P. and Clarke, C.F. (2012) Folate status of gut microbiome
hyperhomocysteinemia and thrombosis: a case and control study. affects Caenorhabditis elegans lifespan. BMC Biol. 10, 66
Int. J. Hematol. 93, 458464 57 Martin, G.M. (2011) The biology of aging: 1985-2010 and beyond.
29 Hoffbrand, A.V. (2012) Megaloblastic anemias. In Harrisons Principles FASEB J. 25, 37563762
of Internal Medicine (18th edn) (Longo, D.L. et al., eds), pp. 862871, 58 Cabreiro, F. et al. (2013) Metformin retards aging in C. elegans by
McGraw-Hill altering microbial folate and methionine metabolism. Cell 153, 228
30 Kumar, N. (2014) Neurologic aspects of cobalamin (B12) deficiency. 239
Handb. Clin. Neurol. 120, 915926 59 Onken, B. and Driscoll, M. (2010) Metformin induces a dietary
31 Pepper, M.R. and Black, M.M. (2011) B12 in fetal development. Semin. restriction-like state and the oxidative stress response to extend C.
Cell Dev. Biol. 22, 619623 elegans healthspan via AMPK, LKB1, and SKN-1. PLoS ONE 5, e8758

502
Downloaded for Tinnaphop Daraswang (tinnaphop8109@gmail.com) at Chulalongkorn University from ClinicalKey.com by Elsevier on May 09, 2017.
For personal use only. No other uses without permission. Copyright 2017. Elsevier Inc. All rights reserved.
Review Trends in Genetics November 2014, Vol. 30, No. 11

60 Martin-Montalvo, A. et al. (2013) Metformin improves healthspan pro-longevity effects of oral supplementation with lipid-soluble
and lifespan in mice. Nat. Commun. 4, http://dx.doi.org/10.1038/ antioxidants. Biogerontology 13, 337344
ncomms3192 69 Bentinger, M. et al. (2010) Coenzyme Q biosynthesis and functions.
61 Pyra, K.A. et al. (2012) Prebiotic fiber increases hepatic acetyl CoA Biochem. Biophys. Res. Commun. 396, 7479
carboxylase phosphorylation and suppresses glucose-dependent 70 Vasta, V. et al. (2011) Altered redox status of coenzyme Q9 reflects
insulinotropic polypeptide secretion more effectively when used with mitochondrial electron transport chain deficiencies in Caenorhabditis
metformin in obese rats. J. Nutr. 142, 213220 elegans. Mitochondrion 11, 136138
62 Shin, N.R. et al. (2014) An increase in the Akkermansia spp. population 71 Jonassen, T. et al. (2002) Development and fertility in Caenorhabditis
induced by metformin treatment improves glucose homeostasis in diet- elegans clk-1 mutants depend upon transport of dietary coenzyme Q8 to
induced obese mice. Gut 63, 727735 mitochondria. J. Biol. Chem. 277, 4502045027
63 Karlsson, F.H. et al. (2013) Gut metagenome in European women with 72 Larsen, P.L. and Clarke, C.F. (2002) Extension of life-span in
normal, impaired and diabetic glucose control. Nature 498, 99103 Caenorhabditis elegans by a diet lacking coenzyme Q. Science 295,
64 Barja, G. (2013) Updating the mitochondrial free radical theory of 120123
aging: an integrated view, key aspects, and confounding concepts. 73 Saiki, R. et al. (2008) Altered bacterial metabolism, not coenzyme Q
Antioxid. Redox Signal. 19, 14201445 content, is responsible for the lifespan extension in Caenorhabditis
65 Ernst, I.M.A. et al. (2013) Vitamin E supplementation and lifespan in elegans fed an Escherichia coli diet lacking coenzyme Q. Aging Cell 7,
model organisms. Ageing Res. Rev. 12, 365375 291304
66 Pallauf, K. et al. (2013) Vitamin C and lifespan in model organisms. 74 Gomez, F. et al. (2012) Delayed accumulation of intestinal coliform
Food Chem. Toxicol. 58, 255263 bacteria enhances life span and stress resistance in Caenorhabditis
67 Shibamura, A. et al. (2009) A method for oral administration of elegans fed respiratory deficient E. coli. BMC Microbiol. 12, http://
hydrophilic substances to Caenorhabditis elegans: effects of oral dx.doi.org/10.1186/1471-2180-12-300
supplementation with antioxidants on the nematode lifespan. Mech. 75 Chin, R.M. et al. (2014) The metabolite a-ketoglutarate extends
Ageing Dev. 130, 652655 lifespan by inhibiting ATP synthase and TOR. Nature 510, 397401
68 Kashima, N. et al. (2012) Development of a method for oral 76 Stanfel, M.N. et al. (2009) The TOR pathway comes of age. Biochim.
administration of hydrophobic substances to Caenorhabditis elegans: Biophys. Acta 1790, 10671074

503
Downloaded for Tinnaphop Daraswang (tinnaphop8109@gmail.com) at Chulalongkorn University from ClinicalKey.com by Elsevier on May 09, 2017.
For personal use only. No other uses without permission. Copyright 2017. Elsevier Inc. All rights reserved.

Anda mungkin juga menyukai