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EVs Flashcards

1
Q

What are extracellular vesicles (EVs)?

A

EVs are membrane-bound vesicles secreted by all cell types involved in intercellular communication.

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2
Q

Why are EVs considered evolutionarily conserved?

A

The secretion of EVs is a fundamental cellular process found across different organisms.

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3
Q

What was the initial purpose of EV secretion?

A

EVs were thought to be a way for cells to eliminate unneeded compounds.

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4
Q

What is the current understanding of EV function?

A

EVs play a crucial role in cell-to-cell communication in both normal and pathological processes.

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5
Q

Are EVs capable of replication?

A

No
EVs do not replicate like viruses.

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6
Q

What makes EVs heterogeneous? EVs vary in size

A

EVs vary in size composition

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7
Q

What are the three main types of EVs? Exosomes

A

Exosomes
microvesicles
apoptotic bodies

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8
Q

What distinguishes exosomes from other EVs?

A

Exosomes originate from multivesicular endosomes whereas microvesicles bud directly from the plasma membrane.

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9
Q

How do EVs contribute to homeostasis?

A

They transport proteins and lipids

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10
Q

Can EVs carry genetic material?

A

yes
EVs can transport DNA

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11
Q

When was the first evidence of EVs found? In 1946

A

In 1946 Erwin Chargaff and Randolph West observed procoagulant properties in ultracentrifuged blood plasma.

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12
Q

When was the term “extracellular vesicle” first used?

A

1971

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13
Q

Which labs contributed to understanding EVs in the 1980s?

A

The Stahl and Johnstone labs studied EV release from reticulocytes and tumor cells.

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14
Q

How were EVs linked to immune responses in the 1990s?

A

Graça Raposo’s lab found that EVs played a role in antigen presentation and immune regulation.

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15
Q

What was discovered about EVs in 2006?

A

Researchers found that EVs contain nucleic acids and can transfer them between cells.

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16
Q

How did early studies analyze EVs?

A

Using ultracentrifugation and electron microscopy

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17
Q

What was the significance of the 1980s studies on tumor cells?

A

They revealed that cancer cells release EVs potentially influencing tumor progression.

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18
Q

What milestone did EV research reach in the 1990s?

A

EVs were recognized for their role in the immune system.

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19
Q

Why is 2006 an important year for EV research?

A

It marked the discovery of EV-mediated nucleic acid transfer.

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20
Q

What is the significance of ultracentrifugation in EV studies?

A

It allows for the isolation and characterization of EV populations.

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21
Q

What are microvesicles?

A

EVs that bud directly from the plasma membrane.

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22
Q

What are exosomes?

A

Small EVs formed inside multivesicular bodies and released when these fuse with the plasma membrane.

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23
Q

What are apoptotic bodies?

A

Large EVs formed during programmed cell death.

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24
Q

What is ESCRT?

A

Endosomal Sorting Complex Required for Transport which helps in exosome formation.

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25
How are exosomes released? By fusion of multivesicular endosomes with the plasma membrane.
By fusion of multivesicular endosomes with the plasma membrane.
26
What distinguishes microvesicles from exosomes? Microvesicles bud directly from the plasma membrane
Microvesicles bud directly from the plasma membrane while exosomes originate from endosomes.
27
What are MDVs?
Mitochondrial-derived vesicles a special class of EVs involved in mitochondrial quality control.
28
What regulates EV biogenesis? Cellular signals
Cellular signals stress conditions
29
What influences EV composition?
The origin of the EV-producing cell and the conditions in which it exists.
30
Do all cells release EVs?
yes EV secretion is a universal cellular process.
31
What are the four possible fates of EVs in recipient cells? Surface binding
Surface binding intracellular signaling content recycling content release to cytoplasm
32
How do EVs participate in antigen presentation?
They can transfer antigens to immune cells triggering immune responses.
33
What role do EVs play in metabolism?
They transport metabolic molecules that can be recycled by recipient cells
34
How can EVs mediate signaling?
By interacting with cell surface receptors.
35
How can EVs influence gene expression?
Through the transfer of RNA molecules.
36
What is one function of mitochondrial-derived vesicles (MDVs)?
They help in mitochondrial quality control.
37
How does oxidative stress affect EV release?
It can trigger the release of EVs containing mitochondrial DNA.
38
What is one way EVs contribute to neuroinflammation?
By activating microglia.
39
How do EVs participate in inter-organ communication?
They carry signaling molecules that affect distant tissues.
40
How can EVs influence cancer progression?
They transfer oncogenic signals that promote tumor growth and metastasis.
41
What are two types of age-related macular degeneration (AMD)?
Dry AMD and wet AMD.
42
What is the role of EVs in AMD?
They may carry harmful mitochondrial DNA contributing to disease progression.
43
How do EVs contribute to oxidative stress in retinal cells?
By releasing mitochondrial DNA.
44
What is one biomarker found in EVs?
mtDNA.
45
What is eNAMPT?
A protein in EVs that enhances NAD+ biosynthesis and affects aging.
46
How does eNAMPT affect aging?
Higher plasma eNAMPT levels correlate with extended lifespan.
47
What is one potential role of EVs in traumatic brain injury (TBI)?
They carry biomarkers that indicate injury severity.
48
How might EVs be used therapeutically?
As drug delivery vehicles.
49
What are brain-derived EV markers?
Proteins and nucleic acids specific to neuronal cells found in EVs.
50
What disease areas are EV biomarkers being studied in?
Cancer neurodegeneration
51
What is ultracentrifugation used for in EV research?
It is a common method for isolating EVs from biological fluids.
52
What is Nanoparticle Tracking Analysis (NTA)?
A technique used to measure the size and concentration of EVs in a sample.
53
How does electron microscopy help in EV studies?
It provides high-resolution images of EV morphology.
54
What is flow cytometry used for in EV analysis?
It helps in characterizing and quantifying EVs based on surface markers.
55
Why is western blotting used in EV research?
To detect specific EV-associated proteins.
56
What is RNA sequencing used for in EV studies?
To analyze the RNA content of EVs and identify biomarkers.
57
What is the significance of proteomics in EV research?
It helps identify the protein composition of EVs.
58
How are EVs labeled for tracking studies?
Using fluorescent dyes or genetic labeling techniques.
59
What is the role of microfluidic devices in EV isolation?
They enable rapid and efficient separation of EVs from biofluids.
60
Why is standardization important in EV research?
To ensure reproducibility and comparability of results across studies.
61
How are EVs being explored for drug delivery?
They can be engineered to carry therapeutic molecules.
62
What is the role of EVs in personalized medicine?
They provide biomarkers that can be used for patient-specific diagnosis and treatment.
63
How are EVs implicated in neurodegenerative diseases?
They carry misfolded proteins that may contribute to disease progression.
64
What is the connection between EVs and cardiovascular diseases?
EVs transport inflammatory and coagulation-related molecules.
65
How do tumor-derived EVs contribute to metastasis?
They transfer oncogenic signals and create a pre-metastatic niche.
66
What is the role of EVs in aging research?
They influence cellular senescence and systemic aging processes.
67
How are EVs studied in the context of infections?
Some pathogens hijack EVs for immune evasion and viral transmission.
68
What are hybrid EVs?
Artificially engineered EVs that combine synthetic and natural components.
69
How are EVs being integrated into liquid biopsy approaches?
They provide a non-invasive way to analyze disease biomarkers.
70
What is one major challenge in EV research?
The heterogeneity of EVs makes standardization and interpretation difficult.
71
examples of EV content
DNA RNA small molecules proteins disease specific cargo tetraspanins cell-specific markers lipid rafts HLAs Integrin
72
current techniques for EV isolation
ultracentrifugation immunomagnetic bead capture ultrafiltration precipitation microfluidics ELISA field flow fractionation affinity purification SEC HPLC
73
considerations in selecting an isolation method
type of sample future experiments yield? purity? time? starting sample volume?
74
considerations with sample type
vesicle concentration? volume? viscosity? complexity?
75
what are some downstream experiments to do on EVs
flow cytometry NTA western blot proteomic analysis RNA analysis functional analysis therapeutics
76
pros and cons of ultracentrifugation
pros - straightforward cons - purity and integrity
77
density centrifugation pros and cons
pros - high purity cons - slow, sucrose toxicity
78
ultrafiltration pros and cons
pros - quick cons - low purity
79
immunomagnetic bead pros and cons
pros - specific, high purity cons - prior knowledge of characteristics required
80
affinity purification pros and cons
pros - relatively specific, high purity cons - prior knowldge of EV characteristics required
81
chromatography pros and cons
pros - high purity cons - equipment
82
microfluidics pros and cons
pros - specific, high purity cons - prior knowldge of characteristics required
83
field flow fractionation pros and cons
pros - quick cons - purity
84
precipitation pros and cons
pros - high yield cons - low purity
85
dry AMD
cellular debris called drusen accumulates between the retina and the choroid, causing damage to the retina and photoreceptors.
86
wet AMD
blood vessels infiltrate from the retina to the choroid and cause hemorrhaging

GSBS Cell Bio (6 decks)

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