INT303326

Context Info Confidence 0.75 First Reported 2010 Last Reported 2010 Negated 1 Speculated 1 Reported most in Body Documents 25 Total Number 26 Disease Relevance 10.08 Pain Relevance 0.76

cytosol (CHMP2B) endosome (CHMP2B) mitochondrion (CHMP2B) nucleus (CHMP2B) intracellular (CHMP2B) cytoplasm (CHMP2B)

Anatomy Link Frequency Neuron 3 HeLa 2 brain 1 vesicles 1 motor neurons 1 CHMP2B (Homo sapiens)

Pain Link Frequency Relevance Heat Spinal cord 175 97.20 Hippocampus 50 95.40 midbrain 50 94.72 medulla 100 90.80 withdrawal 50 90.12 Kinase C 25 53.64 backache 25 29.00 cytokine 25 24.28 carbamazepine 1 5.00 Clonidine 1 5.00

Disease Link Frequency Relevance Heat Dementia 376 100.00 Motor Neuron Diseases 1554 99.68 Frontotemporal Dementia 53 99.52 Death 79 99.32 Repression 75 99.32 Neuroblastoma 25 99.00 Disease 340 95.20 Neurodegenerative Disease 216 94.40 Dysphagia 25 85.20 Dysarthria 50 84.76

Sentences Mentioned In

Key:

Protein

Mutation

Event

Anatomy

Negation

Speculation

Pain term

Disease term

Depletion of ESCRT-III or overexpression of Vps2b mutant proteins in cortical neurons presents with a similar phenotype [52].

Gene_expression (overexpression) of Vps2b in neurons

1) Confidence 0.75 Published 2010 Journal Seminars in Cell & Developmental Biology Section Body Doc Link PMC2938570 Disease Relevance 0.63 Pain Relevance 0

The small vacuoles found in cells expressing WT CHMP2B co-localised with large areas of CD63 staining – displaying the normal physiological fusion of late endosomes with lysosomes (Figure 6 F–H).

Gene_expression (expressing) of CHMP2B

2) Confidence 0.68 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0.18 Pain Relevance 0

In cells expressing WT CHMP2B, CD63 co-localises with small vacuoles within the cytoplasm (Figure 6F–H).

Gene_expression (expressing) of CHMP2B

3) Confidence 0.68 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.04

CHMP2B is expressed in all major areas of the human brain, as well as multiple other tissues outside the CNS.

Gene_expression (expressed) of CHMP2B in brain

4) Confidence 0.68 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 1.15 Pain Relevance 0.07

The number of cells with halos in cells expressing the p.T104N isoform of CHMP2B was significantly increased (Bonferroni post-test, p<0.05).

Gene_expression (expressing) of CHMP2B

5) Confidence 0.68 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.05

Image analysis showed there were significantly more vacuoles with an area greater than 1µm2 in cells expressing mutant CHMP2B protein compared to wild-type (one way ANOVA: p<0.0001, Figure 7).

Gene_expression (expressing) of CHMP2B

6) Confidence 0.68 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.04

In cells expressing mutant CHMP2B, CD63 does not co-localise with the large cytoplasmic vacuoles caused by mutant CHMP2B transfection, but instead is found on the membrane of these aberrant structures (Figure 6 I–K).

Gene_expression (expressing) of CHMP2B

7) Confidence 0.68 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.04

Additionally, there is strong evidence that CHMP2B mutations result in neuronal death; dendritic retraction and cell death seen in cortical neuron cultures transfected with CHMP2BIntron5 is a result of the failure of the aberrant protein to dissociate from the other subunits of the ESCRTIII prior to the next round of sorting, a step critical for the formation of MVBs [12].

Gene_expression (transfected) of CHMP2BIntron5 in neuron associated with death

8) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0.34 Pain Relevance 0

In contrast, cells expressing mutant CHMP2B often showed CD63 immunostaining on the membranes of large vacuoles (Figure 6 I–K).

Gene_expression (expressing) of CHMP2B

9) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0.20 Pain Relevance 0

Transfection of HeLa cells with CHMP2BIntron5 and CHMP2B?

Gene_expression (Transfection) of CHMP2BIntron5 in HeLa

10) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0.30 Pain Relevance 0

Co-staining with CD63, a tetraspanin that is abundant in late endosomes and lysosomes, revealed an interesting change in staining pattern in cells expressing mutant CHMP2B.

Gene_expression (expressing) of CHMP2B

11) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.04

In the present study, we have demonstrated that LC3-II levels are significantly increased in cells expressing mutant CHMP2B.

Gene_expression (expressing) of CHMP2B

12) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0.20 Pain Relevance 0

In cells expressing mutant CHMP2B, CD63 does not co-localise with the large cytoplasmic vacuoles caused by mutant CHMP2B transfection, but instead is found on the membrane of these aberrant structures (Figure 6 I–K).

Gene_expression (transfection) of CHMP2B

13) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.03

Cells expressing mutant CHMP2B isoforms had large cytoplasmic vacuoles that were devoid of lumenal staining for the recombinant protein, such vacuoles were rarely seen in cells expressing wild-type protein (Figure 6B–D).

Gene_expression (expressing) of CHMP2B

14) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.03

Transfection of HeLa cells with CHMP2BIntron5 and CHMP2B?

Gene_expression (Transfection) of CHMP2B in HeLa

15) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0.30 Pain Relevance 0

In vitro models to investigate the functional effects of CHMP2B mutations

Spec (investigate) Gene_expression (mutations) of CHMP2B

16) Confidence 0.59 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0.07 Pain Relevance 0

Transfection of mutant CHMP2B into HEK-293 and COS-7 cells resulted in the formation of large cytoplasmic vacuoles, aberrant lysosomal localisation demonstrated by CD63 staining and impairment of autophagy indicated by increased levels of LC3-II protein.

Gene_expression (Transfection) of CHMP2B

17) Confidence 0.52 Published 2010 Journal PLoS ONE Section Abstract Doc Link PMC2844426 Disease Relevance 1.05 Pain Relevance 0

This hypothesis is further supported by the transfection of FTD-related CHMP2B truncating mutations in neuroblastoma cell lines, which led to the formation of large vesicles, with a morphology in keeping with aberrant endosomes [52].

Gene_expression (transfection) of CHMP2B in vesicles associated with neuroblastoma and dementia

18) Confidence 0.52 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0.25 Pain Relevance 0

Cellular pathology of CHMP2B mutations

Gene_expression (mutations) of CHMP2B

19) Confidence 0.51 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.03

Microarray analysis of cervical motor neurons from CHMP2B cases vs. controls

Gene_expression (cases) of CHMP2B in motor neurons

20) Confidence 0.51 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844426 Disease Relevance 0 Pain Relevance 0.38

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