Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Module 3 > Gene therapy in macrophages > Flashcards

Gene therapy in macrophages Flashcards

1
Q

Why is genetic background in macrophages important in inflammatory disease?

A

common genetic variation has an effect on the gene expression levels in macrophages

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Why are macrophages important in inflammatory diseases?

A

they are plastic cells where external stimuli affects the transcriptome and activation phenotype

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

Give an example of where different external stimuli targeting the same receptor can affect the transcriptome and activation phenotype?

A

TLR4 stimulation at the cell membrane results in AP-1 and NFkB expression- inflammatory cytokiens whereas activation at the endosome results in activation of IRF3 and type 1 cytokines

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What do inflammatory cytokines and type 1 IFNs induce in macrophages?

A

antimicrobial functions; chemotaxis; phagocytosis; tissue reapir factors; metabolic regulatory

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

What are the questions surroudn ing genetic control of inflammatory disease with regards to macrophages?

A

how does germline sequence variation control macrophage function; how does germline sequence variation regulating macrophages cause inflammatory disease?

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

Why is crescentic GN in the rat used as a model in looking at macrophages in inflammatory disease?

A

crescentic GN is macrophage dependet

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What different rat strains used to look at macrophages role in crescentic GN?

A

LEW mouse strain is resistant to GN whereas WKY rat strain is susceptible when stimualted with nephrotoxic serum- what are the genetic determinants of this difference?

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

How can the WKY strain and LEW strain be used to look at genetic control of inflammatory disease?

A

if cross the LEW resistnat strain and the WKY susceptbile strain, you create a heterogenous population with a range of phenotypes- look at the genes influencing this

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What is a QTL?

A

quantitative trait locus is a locus which correlates with variation of a quantitative trait in the phenotype of a population of organisms

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

How are expression QTLs used to look at genetic risk?

A

germline variation results in a difference in mRNA levels due to SNPs resulting in quantitative trait transcript —eQTL is a genomic loci that explains the variation in expression levels of mRNA which alters the phenotype of the cell– changes inflammatory disease

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

What is the difference between cis-eQTLs and trans-eQTLs?

A

cis- SNP affecting the QTL is close to the gene whereas trans- SNP is distal to the gene and both result in changes in the expression levels of the gene

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

What is seen in the macrophages of WKY rats vs LEW rats?

A

WKY rats who increased fusion of macrophages to form multinucleating giant cells whereas LEW macorphages don’t

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

When are fused macrophages seen ?

A

ostecloasts and granulomas

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Why is a model of multinucleated giant cell formation useful in GN?

A

in the nephritic glomerulus of WKY rats, multinucleated giant cells are seen

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

What is the experimental design for looking at the genetics determining multinucleated giant cell formation?

A

cross the WKY and LEW rat strains to mix their DNA and create a broad rang of MGC formation and from there look at hte eQTLs

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

What was found to be a genetic determinant of macrophage gene expression in WKY x LEW population?

A

SNP in chromosome 9 was a master regulator of 190 transcripts

17
Q

What gene family did the SNP in chromosome 9 correspond to?

A

Trem family especially Trem2

18
Q

How could the effect of Trem2 in macrophage gene expression be confirmed?

A

use SiRNA and look at the corresponding expression of different genes after the knock down

19
Q

Who was responsible for creating SiRNA?

A

Fire and Mello (2006 Nobel Prize)

20
Q

Why is SiRNA revolutionary in the field of biomedical science?

A

no longer have to genetically create KO mice strains, can be done in cell lines and can image in real time to see the effects of SiRNA

21
Q

What are siRNAs derived from?

A

double stranded RNAs which bind to an enzyme which cuts it up, on strand is selected and remains bound to a protein- which is directed to specific areas of the mRNA which then calatyses the breakdown of that mRNA

22
Q

What type of network does Trem2 regulate?

A

a trans network

23
Q

What is the function of Trem2 physiologically?

A

bone homeostasis by controlling hte rate of osteoclastogenesis

24
Q

What is the strongest trans-regatuled gene by Trem2?

A

Kcnn4

25
Q

What happened when Kcnn4 is silenced?

A

there is no multineacleated giant cell formation

26
Q

How was the finding that Kcnn4 is implicated in MGC formation further applied after siRNA

A

pharmacological blockade of Kcnn4- TRAM34 gave the same reduction in MGC formation in both human and rat macropahges

27
Q

What indicates that Kcnn4 is implicated in osteoclast formation?

A

when it is knocked out there are far fewer osteoclast numbers per well and this decreases with inceasing pharmacological inhibition with TRAM34

28
Q

Kcnn4 has been implicated in a trans-regulated network involvewd in osteoclast activity and macrophage multinucleation regulated by Trem2, but how does this relate to GN?

A

pharmacological blockade of Kcnn4 reduced susceptibility to glomerulonephritis after nephrotoxic serum injection in WKY rats - 50% reduction in proteinuria

29
Q

How did genetic deletion of Kcnn4 change susceptibility to arthritis?

A

reduced inflammation in collagen induced arthritis; reduced pannus, cartilage damage and bone damage

30
Q

How does Kcnn4 module bone homeostasis?

A

through formation and activation of osteclasts- with increased bone density in KO Kcnn4 mice

31
Q

What is the group that detemrined the role of Kcnn4 in glomerulonephritis; arthritis and bone mass?

A

Kang et al

32
Q

How does CRISPR/Cas9 work?

A

Cas9 and guide RNA target a DNA strand resulting a double strand break whichresults in nonhomologous repair resulting in cleaved DNA is just joined knocking it out or different DNA is added putting mutations at the targeted site resulting in a kock out straight away

33
Q

How was CRISPR/Cas9 used to look at regulators of TNF?

A

Cas9 mouse was transfected with guiding RNA to lots of different genes, knocking them out- the effect on TNFa production was looked at and the guiding RNA sequenced to determine the novel regulatorys

34
Q

How can CRISPR/Cas9 be used to looks at single cell transcriptomes?

A

barcoded beads are given to each cell as well as a specific gRNA, the single cells are then grouped by their guiding RNA and the cells transcriptome is then analysed

35
Q

What is the overall function of CRISPS technology?

A

enables precise and efficient genome editiing in living eukaryotic cells

36
Q

What is the CRISPR system based on?

A

an adaptive immune system used by microbes to defend themselves against invading viruses by recording and targeting their DNA sequences e.g CRISPR locus in E.coli strain matched the sequence of a phage that infected many E.coli strains

37
Q

What is the result of macrophage fusion?

A

leads to the formation of osteoclasts in bone and multinucleate giant cells in chronic inflammatory diseases

38
Q

What have genetic studies in regulation of macrophage multinucleation shown?

A

that germline variation can regulate the multinucleation and influence disease pathogenesis- murine model of TB- genetic looci were associated with production of inflam mediators by macrophages- presence of a heritable genetic component affecting MGC function and susceptibility to granulomatous diseases ; GWAS of MGC-associated diseases TB and GPA showed common variations assocaited with them

39
Q

What is the assocation of loss of function mutations in TREM2 in human bone disease?

A

can cause Nasu-Hakola disease where defective multinucleation in osteoclasts results in impaired bone resorption

Module 3 (33 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions