NCL Research Newsletter June 2016

Content

Main Topic(s):

          6th NCL Research Award (2015)

          Interview with Dr. Grimm - winner of the 6th NCL Research Award

Scientific and Medical Meetings
Medical News & Recent Publications
New Online Resources
Grants, Awards and Open Positions
Miscellaneous 

 

 

Main topic(s)

6th NCL Research Award (2015)

Dr. Christian Grimm (Ludwig-Maximilians University, Munich, Germany) receives the 6th NCL Research Award for his proposal to try and further elucidate the function of CLN3.

The analyses of CLN3 mutants in various organisms and cellular systems points, in particular, at a key role of wildtype CLN3 in homeostasis of the endolysosomal system. Nonetheless, the exact role of the endolysosomal CLN3 transmembrane protein has largely remained an enigma, and reason for addressing this important topic during the 13th National NCL Congress that was organized by the NCL Foundation in Hamburg (Nov 2, 2015). This meeting was entitled “Clinical and translational aspects of CLN3 disease and CLN3 function”. We already briefly reported on this event earlier on our website. Invited presenters covered different topics including clinical MRI in pediatric neurodegenerative diseases

     Dr. Frank Husemann at the 13th National NCL
    Congress

(Dr. Steffi Dreha-Kulaczewski & Prof. Dr. Robert Steinfeld, University Hospital Göttingen, Germany), the value of brain MRI in CLN3 disease (Prof. Dr. Taina Autti, University of Helsinki, Finland), neuroradiological and ophthalmological aspects of CLN3 disease (Dr. Angela Schulz, UKE Hamburg-Eppendorf, Germany), lymphocyte vacuolization and disease severity in CLN3 disease (Dr. Peter van Hasselt, UMC Utrecht, The Netherlands), OCT in NCL mouse models (Dr. Janos Groh, University Hospital Würzburg, Germany). The current state of research into CLN3 biology was reviewed by Dr. Susan Cotman (Massachusetts General Hospital, Boston, USA). Dr. Roberto Iacone (Roche, Basel, Switzerland) reviewed in vitro disease modeling in cardiomyocytes and retinal pigment epithelium, and Prof. Dr. Geert Bultynck (KU Leuven, Belgium) talked about understanding CLN3 function with lessons learned from Bax Inhibitor-1. More specific topics on the role of  lysosomal Ca2+ signalling were covered by Dr. Katja Rietdorf (Open University, Milton Keynes, U.K.) who talked about lysosomal Ca2+ signalling in the heart, Dr. Christian Grimm (Ludwig-Maximilian University, Munich, Germany) who covered mucolipidosis type IV, Ebola, and the function and pharmacology of endolysosomal cation channels, Dr. Diego Medina (TIGEM, Naples, Italy), who talked about lysosomal calcium signalling, autophagy, calcineurin and TFEB, Dr. Claire Mitchell (University of Pennsylvania, Philadelphia, USA), who covered  the role of lysosomes in retinal pigmented epithelial cells and vision loss, and Dr. Sabine Hilfiker (CSIC, Granada, Spain), who talked about altered intracellular calcium handling and endolysosomal trafficking deficits associated with LRRK2-linked Parkinson´s disease. Dr. Lloyd-Evans provided a very interesting working hypothesis for a function of CLN3 that could explain the dysregulation of calcium homeostasis seen within patient cells and the lysosome.

 

Dr. Grimm’s group is specifically interested in the function and therapeutic relevance of ion channels in the endolysosomal system. In particular, he is interested in the role of members of the TRP (transient receptor potential) superfamily that includes TRPML1, TRPML2 and TRPML3. Lysosomes are involved in nutrient sensing and nutrient recycling. The latter occurs through breakdown of proteins, lipids, and other macromolecules by host of enzymes inside the lysosome. Mutations in a wide variety of genes encoding predominantly lysosomal enzymes or lysosomal membrane proteins can cause lysosomal dysfunction and can result in a variety of rare lysosomal storage disorders (LSDs). These include the neuronal ceroid lipofuscinoses (NCLs), the mucolipidoses including mucolipidosis type IV, which is caused by mutations in TRPML1, or the mucopolysaccharidoses. Lysosomal dysfunction is also implicated in common neurodegenerative disorders such as Alzheimer’s disease and Parkinson’s disease, retinal diseases, pigmentation disorders, metabolic diseases, and trace metal (e.g. iron) deficiencies. Highly critical for a proper functioning of the endolysosomal system are fusion and fission processes of organelles within this pathway, and the regulation of ion concentrations within the endolysosomal compartments. Two types of ion channels, the TRPML cation channels (TRPML1, 2 and 3) and the Two-pore channels (TPCs), have recently emerged as important regulators of endolysosomal homeostasis in health and disease. For example, while mutations in TRPML1 cause the lysosomal storage disease mucolipidosis type IV, an activation of TRPML1 by a synthetic ligand rescued neuronal cell phenotypes caused by the loss of FIG4. Defects in this, lipid phosphatase and scaffold protein are the underlying cause of Charcot-Marie-Tooth disease type 4. Lysosomes in CLN3-deficient neurons and FIG4-deficient cells have in common, that they both show increased intra-lysosomal Ca2+ levels. These in turn likely perturb endo-lysosomal function. Healthy channels still present in the membrane of CLN3-deficient lysosomes might therefore be able to compensate for the loss of CLN3 function. Such channels seem likely therapeutic candidates for treating CLN3 and perhaps other lysosomal storage diseases. Last but not least, it should also not a priori be excluded that compounds that can target wildtype CLN3 might have therapeutic potential in some diseases.
To study the physiological roles and activation mechanisms of lysosomal channels, Dr. Grimm’s laboratory combines lysosomal patch-clamp techniques, molecular biology, cell biology, and mouse genetics. Building on his experience in the endolysosomal channel field, Dr. Grimm’s goal is to try and elucidate a potential role and the regulation of CLN3 as an endolysosomal channel.

 We asked Dr. Grimm some questions which he kindly answered:

How did you become aware of CLN3 disease?
 
“I became aware of the disease for the first time when I was invited as a speaker to the NCL conference in Hamburg in 2015”.

What made you decide to apply for the NCL Research Award?

     Dr. Grimm received the 6th NCL Research Award 
     and his Postdoc Dr. Cheng-Chang Chen (left)

 

"During the NCL conference in Hamburg in 2015 interesting novel research results were presented in particular by Dr. Lloyd Evans (Cardiff) that pointed to a potential role for CLN3 as a novel ion channel or transporter in the endolysosomal system. Given our latest successful efforts in characterizing novel endolysosomal cation channels such as the two-pore channels and the mucolipins (TRPML channels) by using the recently established whole-endolysosomal patch-clamp technique combined with small molecule agonists that we had identified in high-throughput screenings, it was just a logical consequence to think that a similar approach might also work out for CLN3. In particular, it was clear that we would need to develop a small molecule tool to characterize potential ionic features of CLN3”.

Your lab is one of the few in the world that master the technique of patch-clamping lysosomes. Can you explain how this technology might help to elucidate the function of the CLN3 protein and its mutant forms?

“Patch-clamp is the gold standard in ion channel research. Organellar patch-clamp has only become available recently and many potential novel ion channels in endosomes and lysosomes await in-depth characterization. Hence, if CLN3 is indeed an ion channel, the patch-clamp technique would be the best way to either confirm or refute this hypothesis. If successful, this will pave the way to a much more complete and detailed understanding of the functional features of this protein”.

The majority of CLN3 patients carry a CLN3 allele that encodes a truncated protein. Reports suggest that this mutant protein gets stuck in the endoplasmic reticulum, and does not reach the lysosome. Therefore, the mind boggling question how can a better understanding of wildtype CLN3 function pave the road to therapy for CLN3 patients?

“Characterizing wildtype CLN3 is one important milestone in understanding the CLN3 protein and consequently NCL disease in more detail. Developing a therapy for those patients who have truncated versions of CLN3 or who have point mutations that lead to a complete mislocalization of the protein is a different goal. One promising approach would be to 1) become aware of the functional features of CLN3 such as the question whether it is an ion channel and if yes whether it is a cation channel, whether it is nonselective etc. and 2) to see whether exactly this function may possibly be replaced by activation other alternative channel proteins in endosomes/lysosomes such as e.g. TRPML channels or two-pore channels. Hence, in parallel to the search for small molecule agonists of CLN3, a more detailed investigation of TRPML channel agonists or the development of small molecule agonists for TPCs will help to evaluate if those channels may be suitable candidates to rescue the effects of a CLN3 loss in NCL patients”.

What other collaborations would you like to see to support your project?

“In particular, we will need collaboration partners to help us evaluate the effects of TRPML, TPC and CLN3 small molecule agonists in CLN3 KO mouse models and in NCL patient-derived cell lines to see whether NCL-related cellular or whole-organism defects can be rescued or improved with these tools”.

 

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Scientific and Medical Meetings

 

Minisymposium on rare diseases, Münster, February 13, 2016

Fellows of the German Study Foundation (“Studienstiftung des deutschen Volkes”) organized a minisymposium on rare diseases, to discuss recent findings, experiences and lessons-learned from research, therapy and patient care. Dr. Herman van der Putten (NCL Foundation, Hamburg) discussed recent preclinical and clinical findings regarding NCLs and CLN3 specifically, Prof. Dr. Robert Steinfeld (University Medical Center Göttingen) covered more broadly preclinical and clinical findings in neurodegenerative diseases, Dr. Holm Zaehres (Max-Planck-Institute for Molecular Biomedicine, Münster) discussed the history of stem cells and their applications in drug discovery and therapy, and Ms Nelly Burauen (Social worker and care giver) gave a moving presentation that provided excellent insights into everyday’s life challenges that care takers and patients face. A long and lively discussion covering different topics closed the meeting.  


 

ProRetina Potsdam Meeting, April 8-9, 2016

 

This year’s ProRetina conference was entitled “RETINAL DEGENERATION. In Times of Transition and Translation”, and took place in Potsdam (April 8-9, 2016). The conference covered a wide range of topics, some of which perhaps also relevant to NCL. Topics and presentations included historical overviews and perspectives on inherited retinal dystrophy and Müller cells, new stem cell-derived models for compound screening, biomarkers, outcome monitoring in clinical trials, as well as some clinical trials.

For the full program please consult here.

 


42nd Annual Conference of the Neuropediatrics Association, April 21-24, 2016, Bochum

A symposium on the topic “palliative care for progressive disease in children” was held at the Annual Conference of the Neuropediatrics Association (“42. Jahrestagung der Gesellschaft für Neuropädiatrie e.V.”). This symposium included talks by Prof. Robert Steinfeld (Göttingen) on diagnostics and therapeutics for neurodegenerative diseases and Dr. Margit Baumann-Köhler (Münster) on ambulant care for children with chronic diseases. In particular, the talk of Anne and Dirk Schultheis raised a high level of attention and positive feedback. These parents lost their daughter Judith, who suffered from JNCL. They focused, from a family perspective, on the experiences and challenges of treatment and care for children suffering from JNCL.


 
Upcoming Conferences
 

June 12-17, 2016, Lisbon, Portugal: FASEB Conference

Calcium and Cell Function


July 7-10, 2016, Clare College, Cambridge, UK: The Pathobiology of the Lysosome and Lysosomal Disease Conference


July 14-17, 2016, St. Louis, USA: BDSRA Annual Conference


July 30-August 4, 2016, Mount Snow, West Dover, Vermont: Organellar Channels & Transporters Gordon Research Conference


August 7-12, 2016, Girona, Spain: Neurobiology of Brain Disorders, Gordon Research Conference


September 16, 2016, Hamburg, Germany: German Pediatrician Association ("Deutsche Gesellschaft für Kinder- und Jugendmedizin e.V.")

A symposium organized by the NCL Foundation on "Diagnostics and therapy of neurodegenerative diseases in childhood” is taking place in Hamburg.

Preliminary programme:

15:30 – 15:50: Prof. Robert Steinfeld (Göttingen): Rational diagnostic and therapy options for childhood dementia disorders

15:50 – 16:10: Dr. Angela Schulz (Hamburg): Enyme replacement therapy for late-infantile NCL (CLN2)

16:10 – 16:30: Dr. Carola Hasan (Datteln): Patients with juvenile NCL – tasks for the palliative care

16:30 – 16:50: Prof. Dr. med. Thomas Klopstock (Munich): Neurodegeneration with Brain Iron Accumulation (NBIA): Clinics, genetics, therapy

16:50 – 17:00: Angelika Klucken (Velbert): NBIA disorder for medical care – family perspective

 

October 5-8, 2016, Boston, USA: The 15th International Conference on Neuronal Ceroid Lipofuscinosis (Batten Disease)

This international conference takes place in Boston and will cover patient registries, clinical trials, biomarkers and clinical readouts, preclinical models, molecular mechanisms and the identification of drug targets and new therapies. The conference is chaired by Dr. Susan Cotman, and co-chair is Katherine Sims, M.D. (both at Massachusetts General Hospital Boston). Amongst the co-organizers is also Dr. Sara Mole (University College London), who coordinates the BATCure Research Consortium that has been awarded an EU Horizon2020 Innovation Grant.

Deadlines:

Abstract submission: June 17, 2016

Registration: August 15 (Early Bird Rate), September 28 (Regular Rate), after September 28 (Late Registration Rate)

 

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Medical News & Recent Publications

 

Highlights

 

CLN2 disease
In March 2016, Biomarin announced positive 48-week results (based on the Hamburg Motor + Language CLN2 rating) from its Phase 1/2 pivotal ERT study for cerliponase alfa (recombinant human tripeptidyl peptidase 1) to treat children with CLN2 disease, which lack the soluble lysosomal enzyme tripeptidyl peptidase-1 (TPP1). The results were presented at the WORLDSymposium 2016 in San Diego by Dr. Angela Schulz, Department of Paediatrics, University Medical Center Hamburg-Eppendorf. Biomarin is the first to develop a potential ground-braking enzyme replacement therapy administered directly into the brain ventricles for CLN2 disease. Earlier preclinical results of ERT in the Dachshund model of CLN2 disease paved the road to the clinic. Similar encouraging preclinical studies using TPP1 gene therapy exist in the dog model, suggesting that both ERT and gene therapy may change the outlook for therapy in CLN2 disease.   

CLN3 disease
A gene therapy approach for CLN3 disease is being persued by Abeona Therapeutics. The Company licensed an AAV-based gene therapy program in juvenile Batten disease from the University of Nebraska Medical Center, where initial and ongoing research on AAV Gene Therapy for CLN3 is led by Tammy Kielian, Ph.D.
 
CLN6 disease
Recently, a Charlotte and Gwenyth Gray-foundation sponsored Phase I/IIa gene transfer clinical trial for variant late infantile Neuronal Ceroid Lipofuscinosis (CLN6 disease) was initiated. The trial is the first human, open label, single dose study that uses a self-complementary AAV9 gene therapy vector carrying the healthy CLN6 gene.

 

Publications

 

CLN1
Chandra G et al., 2015 Oct. (Hum Mol Genet): “Cln1 gene disruption in mice reveals a common pathogenic link between two of the most lethal childhood neurodegenerative lysosomal storage disorders.“ The analysis of brain tissue from CLN1(-/-) mice revealed that the CLN10 gene encoding cathepsin D is overexpressed, but the maturation of cathepsin D is disrupted. This suggests a link between infantile and congenital NCL.

Peng S et al., 2015 Nov. (Ann Clin Transl Neurol): “Suppression of agrin-22 production and synaptic dysfunction in Cln1 (-/-) mice.” The investigation of brain tissue from CLN1(-/-) mice and infantile NCL patients suggests a link between oxidative stress on brain cells and synaptic dysfunction.

Tikka S et al., 2016 Mar. (Neuromolecular Med): “Proteomic Profiling in the Brain of CLN1 Disease Model Reveals Affected Functional Modules.” This study investigated changes in brain tissue of mouse models in different stages of CLN1 disease. The results seem to point at a misregulation of processes in mitochondrial cell death, RhoA/Huntington's disease signaling and myelin sheath breakdown.
 
Henderson MX et al., 2016 Apr. (Acta Neuropathol): “Neuronal ceroid lipofuscinosis with DNAJC5/CSPα mutation has PPT1 pathology and exhibit aberrant protein palmitoylation.”
Investigation of DNAJC/CSPα brains has shown that CSPα patients show PPT1 accumulation and miss-localization, but reduced function. It was demonstrated that CSPα is depalmitoylated by PPT1, suggesting that CSPα is the substrate of PPT1.


CLN2
Katz ML et al., 2015 Nov. (Sci Transl Med): “AAV gene transfer delays disease onset in a TPP1-deficient canine model of the late infantile form of Batten disease.”
This study shows a clinical benefit in canine model from rAAV gene transfer of TPP1. High expression was observed in ependymal cells and immunostaining confirmed that recombinant protein is present in brain and spinal cord.

Dyke JP et al., 2016 Jan. (AJNR Am J Neuroradiol): “Brain Region-Specific Degeneration with Disease Progression in Late Infantile Neuronal Ceroid Lipofuscinosis (CLN2 Disease).”
High-resolution MT imaging datasets from CLN2 patients and retrospective cohort of healthy children show three distinct patterns in affect brain regions. The study indicates that brain degeneration in CLN2 patients occur at different rates.


CLN3
Ragbeer SN et al., 2016 Mar. (J Child Neurol): “Remote Assessment of Cognitive Function in Juvenile Neuronal Ceroid Lipofuscinosis (Batten disease): A Pilot Study of Feasibility and Reliability.”
The study involved three diseased children and one healthy sibling using a standardized cognitive assessment test. Results indicate a high agreement between an in-person and a remote evaluator comparing cognitive test scores.

Groh J et al., 2016 May (Glia): “Sialoadhesin promotes neuroinflammation-related disease progression in two mouse models of CLN disease.” Regulation of inflammation-related cell adhesion molecule (Sialoadhesin) in NCL disease contributes to axonal perturbation. The study gives new insights on pathogenesis of NCL and treatment via immune-regulatory strategies.
 
CLN5
Amorim IS et al., 2015 Oct. (Brain Behave):  “Molecular neuropathology of the synapse in sheep with CLN5 Batten disease.” The study shows that mechanisms of previously identified regulators of synaptic pathophysiology in mouse models have similar pathology in large animal models. This represents a possible model to test therapeutics.


CLN6 (clinical trial)

“Phase I/IIa Gene Transfer Clinical Trial for Variant Late Infantile Neuronal Ceroid Lipofuscinosis, Delivering the CLN6 Gene by Self-Complementary AAV9”
This Phase I/IIa study is the first human, open-label, single dose study of self-complementary AAV9 to deliver CLN6 gene. In this safety/efficacy study six patients will be enrolled. Two kids have received the therapy via intrathecal injection already.

Zhang et al., 2016 June (Nat Commun): “Brain-specific Crmp2 deletion leads to neuronal development deficits and behavioural impairments in mice.” Zhang et al. recently reported that brain-specific deletion of Crmp2 leads to neuronal developmental deficits. These findings are perhaps relevant for CLN6 disease. The Weimer laboratory, working on CLN6, has put forward the hypothesis that CLN6 may provide a unique mechanism for localised CRMP-2 signaling. Also of note, at least part of the neuroprotective effects of the cell pentrating ester (LKE), an ester of the neural metabolite lanthionine ketimine (LK), have been ascribed to CRMP-2 (Hensley et al., 2010; Nada et al., 2012).
 
CLN7
Faller KM et al., 2016 Apr. (J Neurosci Res): “The Chihuahua dog: A new animal model for neuronal ceroid lipofuscinosis CLN7 disease?” The study of two littermate Chihuahua dogs revealed the identification of variants in MFSD8 encoding gene CLN7. The clinical and histological similarities to the human form of the CLN7 disease suggest the Chihuahua dog as a new model for the study of CLN7.


CLN10
Ketscher A et al., 2016 Mar. (Biochimie): “Neuroectoderm-specific deletion of cathepsin D in mice models human inherited neuronal ceroid lipofuscinosis type 10.” This study demonstrates that CLN10 is initiated by cells of neuroectodermal origin. A novel cathepsin D allele was generated to investigate tissue-specific functions.


Biomarker
Hersrud SL et al., 2016 Feb. (FEBS J): “Plasma biomarkers for neuronal ceroid lipofuscinosis.”
This study identified seven biomarkers in blood plasma from NCL patients by comparing protein levels that significantly differ from control patients. Biomarkers could be used for testing of therapeutic responses in NCL patients.


Review
Geraets RD et al., 2016 Apr. (Orphanet J Rare Dis): “Moving towards effective therapeutic strategies for Neuronal Ceroid Lipofuscinosis.”
This review provides information on current therapeutic approaches that are tested in preclinical and clinical trials. Additional therapeutic approaches to treat NCLs that are currently in development are being discussed.

Aungaroon G et al., 2016 Apr. (Pediatr Neurol): “Correlation Among Genotype, Phenotype, and Histology in Neuronal Ceroid Lipofuscinoses: An Individual Patient Data Meta-Analysis.”
This meta-analysis included 440 individuals diagnosed with NCL. Genetic testing was done and identification of pathogenic mutations revealed a significant clustering of genotypes into juvenile-onset (CLN3) and infantile-onset phenotypes. For CLN6 genotype, two different clusters of age of onset were recognized, but mostly the adult-onset phenotype. This suggests that NCL genotypes differ in terms of age of onset and phenotype correlation.

 

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New Online Resources

 

Slide Deck: Available Research Tools

 

This tool for researchers is provided by the NCL Foundation, and offers a collection of potentially useful information on NCL for researchers and physicians, new to the NCL field. CLN3 disease is the main topic and the file includes information on CLN3 disease (clinical features, gene and gene defects, prevalence etc.), the CLN3 Protein (expression, localization, function), preclinical CLN3 models, and therapeutic approaches. Other useful links are given, e.g. to an educational movie, several webinars accessible via the Beyond Batten Disease Foundation (US) website, and available tool databases (models, brain banks).

  
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Grants, Awards and Open Positions

 

Call for pilot proposal in childhood neurodegeneration and dementia: 7th NCL Research Award


The NCL Foundation announces and solicits applications for its next NCL Research Award (50,000 euros) to help find a cure for childhood dementia. The aim of this award is to seed-fund outstanding pilot research projects, that hold promise to be sustainable, and aim at providing breakthroughs in CLN3 research. Projects proposed, either at the bench or in the clinic, should hold promise for making a significant contribution towards finding or exploring a therapy. The award is intended to support postdoctoral level scientists. We particularly encourage junior principle investigators and collaborative efforts between clinicians and researchers in academia or industry.   


The application form is available on our homepage:
http://www.ncl-stiftung.de/main/pages/index/p/538
 
Deadline for handing in all required documents: October 31, 2016
  

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Miscellaneous


"Bad Jumper Competition" Fundraising

 

Kim Wager from Oxford PharmaGenesis in UK organized various fundraisers including a “Bad Jumper Competition”. His goal was to raise funds for the NCL Foundation to support NCL research. With help of his colleagues and several supporters, he collected an amount of £2141.45 (€2805,95). THANK YOU!

Kim Wager has completed his PhD at the laboratory of Claire Russell focusing on the NCL zebra fish model and continued as a postdoctoral research fellow at the laboratory of Emyr Lloyd-Evans in Cardiff.

 

                              Kim Wagner and his colleagues from Oxford PharmaGenesis

 

 

Scientific Advisory Board - New Members


The NCL Foundation is pleased to announce and welcome two new members as part of the Scientific Advisory Board.

Angela Schulz, Ph.D., M.D., leads the NCL-consultation center as part of the University Medical Center Hamburg-Eppendorf (UKE) where she works as a neuro-pediatrician. She is the coordinator of the international NCL patient registry (part of the European DEM-Child project) and a member of the BATcure program, a second EU-sponsored program that supports finding a cure for Batten disease. She is the principal investigator in an ongoing clinical study that studies safety, tolerability and efficacy of intracerebroventricular BMN 190 as enzyme replacement therapy for the treatment of CLN2 disease.
Prof. Dirk Isbrandt, Ph.D, M.D., focuses on “channelopathies”, which are caused by disease-associated mutations in ion channels. It includes testing pharmacological treatments to prevent, attenuate, or normalize changes in neural network activity and cognitive performance that are associated with neurological conditions associated with channelopathies. He works at the DZNE (German Center for Neurodegenerative Diseases in Bonn/University of Cologne) for Experimental Neurophysiology.

See also the NCL website: http://www.ncl-stiftung.de/main/pages/index/p/570

 

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