Strengths, limitations & further study Flashcards
(6 cards)
What are the limitations?
- lack of repeats (cohort, brain, vessel level)
- lack of clinical information
- anitbody required subjective identification of VSMCs (localition, morphology) & senescent appearing nuclei (SAHFs)
- PMI (impact tissue/protein denaturation, enzyme activity, pH change)
- Nature of post-mortem analysis (end stage disease, vascular reorganisation starts in 4th decade)
- Low case fatality of cSVD
What were the strengths?
Cohort:
- inclusion criteria
- good matching of cases vs controls
- from two smaller, independent cohorts
Light microscopy:
-allows you to visualise vessel, surrounding tissue, assess morphology but also location & preservation fo tissue
Antibody:
- one of the most studied histone modifications
- previous use for chromatin immunoprecipitation on human tissue
Describe the previous use of your antibody?
1) Miao et al., 2011
- DMT1 susceptibility genes - hypothesis that post-translation histone modifications were present, H3K9me3 was not
2) Turcan et al., 2012
- IDH1 mutation in glioma prognosis, H3K9me3 WAS found
3) Kloth et al., 2012
- Tumour suppressor gene hypermethylation in prostate cancer, associated with worse clinical features
How would you further your study?
GOING TO AMERICA TO DO THIS:
Improve limitations
- more repeats at cohort & brain level
- more clinical information e.g. treatment, quantify hypertension
- antibody: check on human colon, other methods of looking at senescence (beta gal, CDK inhibitors, SASP, heterochromatin protein 1, dna dyes)
- double stain with hsMM (fluorescence)
Further concepts:
- compare to cognitive impairment
- sclerotic index (compared AT VESSEL LEVEL)
- impact on surrounding cells/NVU
Answer unanswered questions:
- if not senescence then what?
- what is the significance of the surviving non-senescent VSMCs?
How do you wish the field of senescence to progress?
- better understanding IN VIVO
- cause or consequence of ageing?
- role in both normal ageing and disease?
How do you wish the field of cSVD to progress?
- better definition & diagnostic criteria to allow better comparison between studies
- many discrepancies of nomenclature
- relationship with co-existing pathology
- neuroimaging to assess vessles IN VIVO
- until then further clues from: hereditary cSVD, peripheral SVD (retina, kidney), current neuroimaging & histological techniques
