Towards
the in vitro understanding of PCDH19-epilesy
Borghi
R.1,2,
Magliocca V.1,2,
Petrini S.3,
Zanni G.1,
Tartaglia M.4,
Bertini E.1,
Moreno S.2,
Compagnucci
C.1,
4
1
Dept. of Neuroscience, Lab. of Molecular Medicine IRCCS, Children’s
Research Hospital Bambino Gesù, Rome, Italy
2
Dept. of Science-LIME, University Roma Tre, Rome, Italy
3Confocal
Microscopy Core Facility, Research Laboratories, Bambino Gesù
Children's Hospital, Rome, Italy
4
Genetics and Rare Diseases Research Division, Children’s Research
Hospital Bambino Gesù, Rome, Italy
Introduction.
PCDH19 (Protocadherin 19), a member of the cadherin superfamily, is
involved in the pathogenic mechanism of an X-linked model of
neurological disease. The biological function of PCHD19 in human
neurons and during neurogenesis is poorly known. Therefore, we
decided to use the model of the induced pluripotent stem cells
(iPSCs) to characterize the molecular and cellular phenotype of
PCDH19 mutated neurons derived from patients’ iPSCs. Previous data
(Compagnucci et al., 2015) show that PCDH19 is expressed in
pluripotent cells, before differentiation, in a homogeneous pattern,
despite its localization is often limited to one pole of the cell.
During neuronal differentiation, positional information on the
progenitor cells assumes an important role in acquiring polarization.
The proper control of the cell orientation ensures a fine balancing
between symmetric (giving rise to two progenitor sister cells) versus
asymmetric (giving rise to one progenitor cell and one newborn
neuron) division. This process results in the polar organization of
the neural tube with a lumen indicating the basal part of the
polarized neuronal progenitor cell; in the iPSC model the cells are
organized in the ‘neural rosette’ and interestingly, PCDH19 is
located at the center of the rosette, with other well-known markers
of the lumen (N-cadherin and ZO-1).
Results.
The data obtained suggest us to pursue the investigation of the plane
of cellular division in neuronal precursors at the stage of the
neural rosette. With this aim we used iPSCs derived from patients’
fibroblasts obtained from skin biopsies. Importantly, we found
alterations in the plane of division of neuronal precursors in the
cell cultures derived from patients’ iPSCs. The observed disruption
of the plane of cell division possibly accounts for the accelerated
rate of neuronal differentiation observed in the patients’ neuronal
cultures. In order to test the effect of PCDH19 reduction on cell
division, we silenced PCDH19 in control iPSCs (derived from a healthy
individual) and analyzed the percentage of altered dividing cells
(with centrosome hyper-amplification). To confirm an effect of
mutated PCDH19 protein on dividing cells, we focused on the dividing
cells at the stage of proliferating iPSCs finding numerous cells with
centrosome hyperamplification in iPSCs silenced for PCDH19.
Discussion.
Our data support the use of iPSCs for in vitro modeling of PCDH19
syndrome, highlighting that PCDH19 has a role in instructing the
apico-basal polarity of the progenitor cells, thus regulating the
development of a properly organized neuronal network. The alterations
in the cell division observed in cells silenced for PCDH19 and in
PCDH19-mutated patients may account for an altered control of the
symmetric versus asymmetric cell division.
PCDH19:
Proteolytic processing and gene regulatory functions
Sylvia
A Newbold, James Wilding, Isabel
Martinez-Garay
Division
of Neuroscience, School of Biosciences, Cardiff University
Mutations
in the X-linked gene PCDH19 lead to epilepsy with cognitive
impairment in heterozygous females. Although the gene codes for a
cell adhesion protein of the cadherin superfamily expected to
localize at the cell membrane, recent reports have implicated PCDH19
in the regulation of gene expression and have identified the protein
in the nucleus.
Our
aim is to determine if PCDH19 undergoes proteolytic cleavage
resulting in the generation of an intracellular fragment (PCDH19-ICD)
that translocates to the nucleus to regulate gene expression. In
addition, we want to identify which genes are regulated by PCDH19 in
neural progenitors and neurons.
To
test this hypothesis, we have analysed how the absence, inactivation
or overexpression of specific proteases affects the generation of
PCDH19-ICD in heterologous cells. We have also generated neurons from
embryonic stem cells to analyze PCDH19 processing in neurons,
including the possible role of neuronal activity. To determine
potential transcriptional targets of PCDH19, we have created a mouse
embryonic stem cell line that overexpresses the cytoplasmic domain of
PCDH19 from the Rosa26 locus (CYTO), and an isogenic PCDH19-knockout
line (KO). This is allowing us to compare the transcriptional
profiles of these embryonic stem cell-derived neuronal progenitors
and neurons in the presence or absence of PCDH19, as well as under
conditions of PCDH19-ICD overexpression.
We
believe that PCDH19 is capable of transducing information about
events happening at the membrane to the nucleus to elicit appropriate
cellular responses and expect that our RNAseq analysis will provide
new mechanistic insights into the function of PCDH19 both during
neurogenesis and in post-mitotic neurons.
INTEGRATED
IN SILICO AND EXPERIMENTAL ASSESSMENT OF DISEASE-RELEVANCE OF
PROTOCADHERIN
19 (PCDH19) MISSENSE VARIANTS
Duyen
H Pham1,2,
Melissa R Pitman4,
Renee Schulz1,
Alison Gardner1,
Sarah E Heron1,2,
Mark A Corbett1,2,
Kavitha Kothur8,9,
Deepak Gill8,9,
Sulekha Rajagopalan11,
Kristy Kolc1,
Benjamin J Halliday10,
Stephen P.Robertson10,
Brigid Regan12,
Heidi E Kirsch14,
Samuel F Berkovic12,
Ingrid E Scheffer12,13,
Stuart M. Pitson4,1,3,
Slave Petrovski5,6,
Jozef Gecz1,2,3,7
1
Adelaide Medical School, The University of Adelaide, Adelaide 5000,
Australia.
2
Robinson Research Institute, The University of Adelaide, Adelaide
5006, Australia.
3
School of Biological Sciences, The University of Adelaide, Adelaide
5000, Australia.
4
Centre for Cancer Biology, University of South Australia and SA
Pathology, Adelaide, South Australia, Australia.
5
Centre for Genomics Research, Precision Medicine and Genomics, IMED
Biotech Unit, AstraZeneca, Cambridge, UK.
6
Department of Medicine, University of Melbourne, Parkville, VIC,
Australia.
7
South Australian Health and Medical Research Institute, Adelaide
5000, Australia.
8
Kids Neuroscience Centre, The University of Sydney.
9 TY
Nelson Department of Neurology and Neurosurgery, The Children’s
Hospital at Westmead, Sydney, NSW 2145, Australia.
10
Department of Women's and Children's Health, Dunedin School of
Medicine, University of Otago, Dunedin 9016, New Zealand.
11
Department of Clinical Genetics, Liverpool Hospital, Liverpool, NSW
1871, Australia.
12
Departments of Medicine, Austin Health, The University of Melbourne,
Melbourne, VIC Australia.
13
Department of Paediatrics, The University of Melbourne, Royal
Children’s Hospital, Melbourne, Florey and Murdoch Institutes,
Melbourne, Australia.
14
Department of Neurology, University of California, San Francisco, CA
94122, USA.
Background:
Assessment of variants currently classified as variants of unknown
significance (VOUS) in genes known to be associated with disease is
an increasingly important task, particularly where precision medicine
trials are underway or planned. However, determining the clinical
relevance of these variants can be challenging, particularly when
functional data for the affected protein is scarce. While the
clinical interpretation of DNA variation has improved significantly
through the implementation of high international standards in the
field, there are still numerous VOUS, which need ongoing and often
very specialized assessment. Pathogenic variants in PCDH19 coding for
the cell adhesion/estrogen receptor transcription coregulatory
protocadherin 19 cause girls clustering epilepsy (PCDH19-GCE).
PCDH19-GCE is an early-onset neurodevelopmental disorder (NDD) of
drug-resistant epilepsy, intellectual disability, autism spectrum
disorder and other neuropsychiatric disturbances. Here we aimed to
systematically assess reported and novel missense variants in PCDH19.
Methods:
We evaluated the performance of 16 selected publicly available in
silico prediction tools including MutPred2
(URL:http://mutpred.mutdb.org/) and ANNOVAR tool-package (URL:
http://wannovar.wglab.org/) to assess the pathogenicity of 322 PCDH19
missense variants selected from two datasets. The first dataset
included 238 PCDH19 missense variants selected from the general
population (female only) gnomAD database
(URL:http://gnomad.broadinstitute.org/). The second dataset included
84 reported PCDH19-GCE disease-causing missense variants. We then
performed integrative in silico (topthree bioinformatics tools and
protein structure modelling) and experimental evaluation (using in
vitro reporter assays) together with InterVar (according to 2015
American College of Medical Genetics and Genomics and the Association
for Molecular Pathology guidelines) to interpret the clinical
significance of 45 PCDH19 variants.
Results:
MutPred2, MutationAssessor and GPP were the top-three performing in
silico tools alone or in combination. The prediction accuracy was
100% for published PCDH19-GCE variants and 78% for population
variants (gnomAD allele frequency >1x10-4). Fifty-six percent of
VOUS (gnomAD allele frequency ≤1x10-4) tested were classified as
deleterious. The assessment toolbox was validated on a set of
recently published, novel PCDH19 variants with high reliability.
Further testing and validation of the toolbox for novel unpublished
PCDH19 variants and also for variants identified in other genes
involved in NDDs such as, TBLXR1 and PCDH12 demonstrated a high
utility and accuracy.
Conclusion:
We have developed a toolbox for the accurate assessment of PCDH19
variant pathogenicity. This is the crucial next step in readiness for
precision medicine to address the severe seizure and behavioural
disorders that occur in females and mosaic males with PCDH19 disease.
Cortical
excitability in a conditional model of PCDH19 Epilepsy
Didi
Lamers1,
Roberta Mezzena1,
Silvia Landi2
, Maria Passafaro3,
Silvia Bassani3
, Gian Michele Ratto1
1
NEST, Scuola Normale Superiore and Instituto Nanoscienze Consiglio
Nazionale delle Ricerche (CNR), Pisa, Italy.
2
Institute of Neuroscience, Pisa, Italy.
3
Institute of Neuroscience, Milan, Italy
PCDH19
Epilepsy is characterized by epileptic seizure onset in early infancy
and is frequently associated with intellectual disability and autism
spectrum disorder1,2,3. The disease has been attributed to mutations
in the X chromosomal Protocadherin 19 (PCDH19) gene1, encoding a
Ca2+-dependent cell-cell adhesion protein. Interestingly, only
heterozygous females and males with somatic mutations are affected4,
leading to the hypothesis that the disease is caused by a mosaic
PCDH19 expression in the brain5. However, the mechanism by which
PCDH19 mosaicism causes epilepsy and cognitive impairment is unknown.
Here,
we employed a novel mouse model of PCDH19 Epilepsy and performed in
vivo electrophysiological and imaging studies. We obtained a focal
mosaic loss of PCDH19 by the injection of an AAV vector carrying
CRE-EGFP in the visual cortex. The opposite hemisphere served as
internal control in the electrophysiological experiments. Local field
potential recordings in anesthetized animals demonstrate that mosaic
PCDH19 patches in the brain have disrupted slow wave activity and
show transient episodes of hyperexcitability and hypersynchronous
activity. Intriguingly, in-depth analysis of the slope of slow wave
activity and single unit statistics, suggest that the local network
has an overall reduced firing rate. These findings are consistent
with the attenuation of slow wave activity but are at odds with the
onset of transient episodes of hyperexcitability.
To
further explore the neuronal activity underlying the observed
phenotype, we performed combined LFP recordings and two photon
calcium imaging of PCDH19 positive and negative neurons in a mosaic
brain in vivo. Preliminary data suggest that some mosaic animals
displayed an increased calcium activity likely to be associated to
the transient hyperexcitability. However, more data are needed to
confirm this indication. No difference was observed between PCDH19
positive and negative neurons in the mosaic animals.
Given
the importance of slow wave activity for both the homeostatic and
memory functions of sleep, and observations that PCDH19 patients
demonstrate abnormal sleep patterns6, our data could be relevant to
patients to increase understanding of the underlying mechanisms of
sleep disturbances in PCDH19 Epilepsy.
References
1. Dibbens,
L. M. et al. X-linked
protocadherin 19 mutations cause female-limited epilepsy and
cognitive impairment. Nat. Genet. 40, 776–781 (2008).
2. Marini,
C. et al. Protocadherin 19 mutations in girls with infantile-onset
epilepsy. Neurology 75, 646–53 (2010).
3. Scheffer,
I. E. et al. Epilepsy and mental retardation limited to females: An
under-recognized disorder. Brain 131, 918–927 (2008).
4. de
Lange, I. M. et al. Male
patients affected by mosaic PCDH19 mutations: five new cases.
Neurogenetics 18, 147–153 (2017).
5. Depienne,
C. et al. Sporadic infantile epileptic encephalopathy caused by
mutations in PCDH19 resembles dravet syndrome but mainly affects
females. PLoS Genet. 5, e1000381 (2009).
6. Smith,
L. et al. PCDH19-related epilepsy is associated with a broad
neurodevelopmental spectrum. Epilepsia 59, 679–689 (2018).
Genetic
silencing of PCDH19 as treatment for Early Infantile Epileptic
Encephalopathy type 9 (EIEE-9)
Giorgia
Giansante1,
Elisa Giorgio2,
Marta Ferrero2,
Elisa Pozzi2,
Luca Murru1,
Silvia Bassani1,
Alfredo Brusco2,
Maria Passafaro1
1
Institute of Neuroscience, National Research Council (CNR), Milan
20129, Italy
2
University of Torino, Dept. Medical Sciences, Torino 10126, Italy
Early
Infantile Epileptic Encephalopathy type 9 (EIEE-9) is a form of
epilepsy characterized by a spectrum of neurodevelopmental disorders
with autistic features. Mutations in PCDH19 gene cause a loss of
homophilic cell-cell interactions (Hayashi et
al.,
2017), axon guidance and dendrite self-avoidance (Pederick et
al.,
2018). The proposed pathogenic mechanism is called “cellular
interference”, which is generated by a mosaic expression of PCDH19
in brain cells. In fact, EIEE-9 mainly affects females with
heterozygous PCDH19 mutations and the identification of some male
patients with mosaics expression of PCDH19 strongly supports the
cellular interference hypothesis. In this study we investigated
whether PCDH19 mosaic expression is a critical determinant of altered
neuronal activity in cultured neurons and in acute brain slices.
Moreover, we evaluated if genetic rescue of cellular interference
mediated by RNA interference might represent a promising therapeutic
option for EIEE-9.
Methods:
In order to obtain different levels of PCDH19 mosaic expression in
vitro,
we infected primary cortical and hippocampal neurons obtained from
PCDH19 floxed mice at postnatal day (P)0 with increasing amount of
adenoviral particles expressing the Cre recombinase (AAV-Cre). The
extent of PCDH19 mosaicism has been evaluated by immunocytochemistry
and western blot. Subsequently, we studied extracellular spontaneous
activity of cultured neuronal at two weeks in
vitro. Neurons
infected with a multiplicity of infection chosen to reproduce the
50:50 mosaic condition in
vitro were
analyzed by Micro-Electrode Array (MEA) device. In parallel, MEA has
been used to record spontaneous electrical activity in acute cortical
hippocampal slices from PCDH19 conditional KO mice.
With
the aim of identifying genetic modulators of PCDH19, we have designed
and tested three different siRNAs targeting both the human and the
murine PCDH19 gene in human
and
murine cell lines. Cells have been transfected with selected PCDH19
siRNAs, a siRNA against GAPDH (C+) and a scramble siRNA (C-) at 40nM
and 100nM concentrations. Forty-eight hours post transfection cells
have been harvested and splitted to extract RNA and proteins. PCDH19
levels were evaluated by real-time PCR and western blot analysis.
Results:
Preliminary
results suggested an increase in the duration of the mean burst
activity of cortical neuronal networks in PCDH19 mosaic expression
recorded in spontaneous conditions respect to cortical neurons
expressing PCDH19. Furthermore, we observed an increase in the main
firing and burst parameters of the network recorded in
cortico-hippocampal slices from PCDH19 conditional KO mice compared
to wild-type littermates.
Finally,
we have identified two efficient siRNAs able to efficiently silence
the human and murine PCDH19 gene and to use for future therapeutic
approaches.
Conclusions:
The
preliminary data obtained suggest a possible role of the PCDH19
mosaicism in the functioning of brain circuitry in vitro and
identified two RNA molecules capable of abrogating PCDH19 expression,
opening the way for future therapeutic approaches.
PCDH19
EPILEPSY: preliminary analysis of 55 cases included in RESIDRAS
Ragona
Francesca and Darra
Francesca
on behalf of RESIDRAS Network (N. Specchio, E. Fontana, T. Granata,
R. Guerrini, L. Giordano, M. Nosadini, N. Zamponi, A. Pini, D.
Battaglia, C. Fusco, C. Zucca, M.P. Canevini)
The
Italian Registry of Dravet Syndrome and other syndromes correlated
with genes on SCN1A and PCDH19 currently includes 55 females affected
by PCDH19 epilepsy enrolled by the following Centers: Roma OPBG (15
cases), Verona (9 cases) Milano Besta (8 cases), Firenze Meyer (5
cases), Brescia (5 cases), Padova (3 cases), Ancona (3 cases),
Bologna (2cases2 cases), Roma Gemelli (2 cases) Reggio Emilia (1
case), Bosisio Parini (1 case), Milano S.Paolo (1 case).
The
geographical distribution of patients is the following: Northen Italy
28, Central-South Italy 25, Switzerland 1 and Romania 1.
The
age at last observation ranges from 1 to 27 years (median 10 years);
the present age being from 5 to 32 years (median 13 years).
One
patient is carrying a deletion, in 2 cases the result of genetic
analysis is missing, the remaining 52 patients carry a PCDH19 gene
mutation The age of seizures onset is within the 6th month of life in
10 cases, between 7 and 12 months in 27 cases; between 12 and 24
months in 13 cases and after 2 years of age in 5 patients.
The
age at diagnosis is within 2 years from seizures onset in 22
subjects. In 13 subjects the disease was diagnosed very late (after
the age of 10 years). The first seizure appears in a febrile
condition in 28/55 cases. Seizure semeiology at onset is reported as:
Generalized tonic clonic in 30 cases, Focal motor in 9, Focal non
motor in 3, massive myoclonia in 2 and Undefined Clustered seizures
in 11.
The
psychomotor development at onset is within the normal range in 49
subjects, delayed or clearly impaired in 6 cases. At last follow-up
the seizures persist in 26/50 subjects; respectively in 17/29 of
those aged less than 12 years and 9/21 of the older subjects.
All
subjects but one are taking chronic antiepileptic treatment (13
mono-therapy, 37 polytherapies).
At
last follow-up 15 subjects still have a normal cognitive functioning,
7 patients present a borderline functioning, 28 patients have an
Intellectual Disability of various degree (mild in 11, moderate in 11
and severe in 6 – these data are missing in 5 subjects). Twenty
three patients suffer of behavioral disorders: autistic features in
6, oppositive disorder in 13, attention deficit in 10, impulse
control disorder in 13, obsessive-compulsive disorder in 4.
The
AA will discuss the correlations between the early clinical features
and the global clinical outcome (epilepsy, cognitive and behavioral
outcome).
Targeting
the Gut Microbiota to Treat Epilepsy
Antonio
Leo1,
Emilio Russo1.
1
Department
Science of Health, School of Medicine and Surgery, University of
Catanzaro, Italy.
More
than 65 million people worldwide are affected by epilepsy, “a
heterogeneous group of neurological diseases characterized by an
enduring predisposition to generate spontaneous recurrent seizures
(SRSs) that are often accompanied by neuropsychiatric comorbidities”,
according to International League Against Epilepsy (ILAE) definition
[1]. Accordingly, there is an urgent need for innovative more
effective and better tolerated therapies able to manage epilepsy and
the issues related to it. Regarding to this aim, it is vital to both
better understand the mechanisms underlying seizures onset and to
identify patients at risk to develop epilepsy [2,3]. Very recently,
there has been increasing interest on the role of peripheral inputs
(stimuli) that can interfere with neurodevelopment and be thus
possibly involved in brain diseases such as epilepsy. In tandem,
there is a growing realization of the role of the gut-microbiota, a
complex group of symbiotic microorganisms colonizing the
gastrointestinal tract, in many aspects of brain and behaviour.
Moreover, the microbiota represents one of the major sources of these
peripheral stimuli [4]. Recently, due to the progress in sequencing
technology, it has also been possible to recognize a bidirectional
link between the microbiota and brain. This link is globally named
the microbiota-gut-brain (MGB) axis, including neural, endocrine,
metabolic and immune system/pathways. Interestingly, through this
link, brain and gut produce signals influencing each other to
coordinate functions in health and disease. Therefore, the
gutmicrobiota complex might be involved in brain disorders including
epilepsy by mediating the proexcitatory effect of peripheral
inflammation through immune system activation (e.g. release of
inflammatory mediators), modulating neural networks by production of
neurotransmitters, SCFAs and key dietary amino acids and acting
consequently on the excitation and inhibition (E/I) balance. Based on
this background, a rationale to study the MGB axis and its potential
role in epilepsy exists.
Nowadays,
currently available preclinical and clinical studies, in spite of
some limitations, seem to support the link between microbiota and
epilepsy [5,6]. Regarding to this, a study performed in rats,
reported the effect of a probiotic mixture (Lactobacillus rhamnosus,
Lactobacillus reuteri, and Bifidobacterium infantis) administration
in a pentylenetetrazole (PTZ)-induced model of kindling,
substantively reduces seizure severity and epileptic activity
compared to controls. The intervention had also a positive effect on
the cognitive performances in all experimental groups. At odds, the
concentrations of oxidant factors, nitric oxide and malondialdehyde,
were reduced in the probioticadministered groups. Lastly, elevated
levels of GABA have been found in brain, but only in the group with
probiotics administration during kindling [7]. Regarding clinical
evidence, few comparative and interventional studies have been so far
performed on the link between gut microbiota and epilepsy.
Recently,
it has been demonstrated a clear difference in gut microbiota
composition between drug-resistant and drug-sensitive patients with
epilepsy (PWE). In details, α-diversity was increased and gut
microbiota was enriched in rare microbes in the drug resistant group
(i.e. Verrucomicrobia), whereas bacteria population and α-diversity
was similar between control and drugs-sensitive patients [8].
Accordingly, it could be hypothesized that the gut microbiota and its
mediators could have a role in drug-resistant mechanisms.
Surprisingly, it has also been shown that the ketogenic diet, used in
the pediatric population with drugs-resistant epilepsy, changes the
composition and function of the gut microbiome both in animal model
of epilepsy and in PWE. Likewise, studies in mice model of epilepsy
have demonstrated that the gut microbiota was necessary for the
therapeutic effect of this diet [6]. Regarding the use of pre- or
probiotics in PWE, one study has evaluated the effects of 4-months
probiotic supplementations (a mixture containing 8 different
bacterial subspecies of Lactobacillus, Bacteroides and Streptococcus
species) in 45 patients with drug-resistant epilepsy. The study
revealed that probiotics supplementation decreased seizure number and
improved the Quality of Life score. However, authors did not perform
any microbiota analysis before and after supplementation [9].
Furthermore, doubtful evidence also exist for the association between
seizure incidence and antibiotic treatment [10]. Overall, multiple
and not yet completely understood mechanisms could be involved in
this bidirectional link and, although alterations in the gut
microbiota and the epileptic process have not been directly
investigated, the nature of these connections suggest their potential
significance to epilepsy. Accordingly, several future studies are
mandatory to establish whether microbe-based treatments can be
effectively and securely used for clinical improvement of seizure
frequencies, severity and epilepsy-related disorders. Likewise,
future researches should be aimed at understanding whether the
epilepsy itself shapes first the microbiota, or whether differences
in microbiota-composition (during lifetime) can be the cause of
seizures onset and maintenance.
Elucidating
the relationship between the MGB axis and epilepsy could also lead to
the discovery of innovative therapies as well as useful biomarkers of
illness advancing the knowledge on the complex mechanisms underlying
epileptogenesis and epilepsy themselves.
References
1 Fisher
RS, Cross JH, French JA, Higurashi N, Hirsch E, Jansen FE, et al.
Operational classification of seizure types by the International
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Classification and Terminology. Epilepsia 2017;58:522–30.
doi:10.1111/epi.13670.
2 Pitkanen
A, Loscher W, Vezzani A, Becker AJ, Simonato M, Lukasiuk K, et al.
Advances in the development of biomarkers for epilepsy. Lancet Neurol
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G, Pauletti A, Pascente R, Vezzani A. Preventing epileptogenesis: A
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Res 2016;110:96–100. doi:10.1016/j.phrs.2016.05.009.
4 Cryan
JF, Dinan TG, Bernard C, Pavlov I, Beaumont W, James W, et al.
Mind-altering microorganisms: the impact of the gut microbiota on
brain and behaviour of the nineteenth century through the pioneering
work. Nat Rev Neurosci 2012;13:701–12. doi:10.1038/nrn3346.
5 De
Caro C, Iannone LF, Citraro R, Striano P, De Sarro G, Constanti A, et
al. Can
we “seize” the gut microbiota to treat epilepsy? Neurosci
Biobehav Rev 2019. doi:10.1016/j.neubiorev.2019.10.002.
6 Dahlin
M, Prast-Nielsen S. The gut microbiome and epilepsy. EBioMedicine
2019. 4. doi:10.1016/j.ebiom.2019.05.024.
7 Bagheri
S, Heydari A, Alinaghipour A, Salami M. Effect of probiotic
supplementation on seizure activity and cognitive performance in
PTZ-induced chemical kindling. Epilepsy Behav 2019;95:43–50.
doi:10.1016/j.yebeh.2019.03.038.
8 Peng
A, Qiu X, Lai W, Li W, Zhang L, Zhu X, et al. Altered
composition of the gut microbiome in patients with drug-resistant
epilepsy. Epilepsy Res 2018;147:102–7.
doi:10.1016/j.eplepsyres.2018.09.013.
9 Gomez-Eguilaz
M, Ramon-Trapero JL, Perez-Martinez L, Blanco JR. The
beneficial effect of probiotics as a supplementary treatment in
drug-resistant epilepsy: A pilot study. Benef Microbes 2018;9:875–81.
doi:10.3920/BM2018.0018.
10 Braakman
HMH, van Ingen J. Can epilepsy be treated by antibiotics? J Neurol
2018;265:1934–6. doi:10.1007/s00415-018-8943-3.
