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Dr. J Specific Topics Flashcards

1
Q

How many viruses induce cancer?

A

2 RNA and 5 DNA viruses

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2
Q

Majority of the RNA viruses belong to which family?

A

Retrovirdae Family

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3
Q

How many retroviruses induce human cancer and what are they?

A

Only one retrovirus induces cancer and it is Human T cell Leukemia Virus 1-HTLV-1

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4
Q

Which virus is the only non-retroviral RNA virus that induces cancer in humans?

A

Hepatitis C

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5
Q

What are the 5 DNA viruses that induce cancer in humans?

A 
EBV
HHV8
HBV
HPV
MCV
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6
Q

What are the two RNA viruses that induce cancer in humans?

A

HTLV

HCV

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7
Q

What percent of human cancers worldwide are caused by oncoviruses?

A

12 (Prevelance in US)

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8
Q

T/F

RNA viruses are associated with breast cancer?

A

False

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9
Q

What are the diseases caused by Epstein Barr Virus EBV?

A

Burkitt Lymphoma
Nasopharyngeal carcinoma
Hodgkin disease
Gastric Carcinoma

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10
Q

What are the disease caused by HHV8?

A

Kaposi sarcoma
Multicentric Castleman disease
Primary Effucsion lymphoma

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11
Q

What are the diseases caused by Human Papilomavirus HPV16 and HPV18?

A

Cervical Carcinoma
Other anogenital carcinomas
Oropharyngeal carcinoma
Non-melanoma skin cancer

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12
Q

What are the diseases cause by Merkel Call Polyomavirus MCV?

A

Merkel Cell Carcinoma

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13
Q

What are the diseases caused by Hepatitis B Virus HBV or Hepatitis C virus HCV?

A

Hepatocellular carcinoma

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14
Q

What are the diseases caused by T-lymphotropic retrovirus HTLV?

A

Adult T cell leukemia/lymphoma

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15
Q

T/F

Oncoviruses are necessary but not sufficient for human cancer development

A

True

This means that viral infection does not always cause cancer. the percentages are low even though viral infections high

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16
Q

T/F Viral cancers are associated with persistent infection and occur immediately after acute infection

A

False

Occur many years after acute infection

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17
Q

What are the two things involved in cancer development?

A

Environment and host co-factors (immunosuppression, genetic predisposition or mutagens)

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18
Q

What can play a deleterious or protective role with some human virus- associated cancers increasing with immunosuppression and other appearing in the context of chronic inflammation?

A

Immune System

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19
Q

What is the first step before viral tumorigenesis?

A

Transformation (In vitro)

This is a development of immortal cells in cell culture

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20
Q

What is oncogenesis?

A

development of tumor in animals/humans

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21
Q

T/F

All transformed cells are oncogenic

A

False

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22
Q

What are the features of transformed cells?

A
  • majority tumorigenic (produce cancer in host)
  • immortal,
  • loss of contact inhibition When they come into contact/touch each other on the plate they will NOT stop dividing, producing a very dense spot of cells on the plate.
  • anchorage independent, meaning they can break off and go into solution or semi-soluble agar.
  • Most are growth factor independent or have a reduced dependence on growth factor.
  • Despite the fact that they may be nutrient deprived, they can still synthesize their DNA
  • aerobic glycolysis Transformed cell still have a lot of oxygen, but they switch from oxidative respiration to glycolysis. This is specific to cancer cells and transformed cell, they start to use glycolysis.
  • Decreased expression of fibers
  • Reduce adhesion to solid substance
  • Produce more surface proteases which allows them to invade other tissues.
  • so you see a redistribution of their microfilaments, which causes the change in shape, - You also see significant change in cell phenotypes. - Acidification of culture medium.
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23
Q

What are the characteristics of normal cells?

A
  • An isolated normal non-tumorigenic cell taken from one organism and introduced to another will not produce a tumor.
  • -They have a finite life span, they are not immortal. - They also have contact inhibition, which means when they grow on a plate, they can’t grow on top of each other. Contact with another cell inhibits this, thus the term, contact inhibition.
  • They are anchorage dependent, if they aren’t blood cells they require a surface to attach to. Loss of that attachment kills them. This is a protective mechanism for cells in our body.
  • They have normal nutrient transfer and normal oxidative respiration to produce ATP–Growth factor dependent-can only grow in culture with growth factor .
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24
Q

Hoe many proteins doe the vast majority of DNA tumor viruses encode for?

A

One oncoprotein that targets host cell factor

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25
Q

What are two of the best known host cell factors targeted by viral proteins?

A

p53 and Rb family members

Bothe act as tumor suppressors during the cell cycle

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26
Q

T/F

Viruses can persist in host cells as part of the episome after integration

A

True

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27
Q

What the activating cell cycle proteins that are targeted by viral oncoprotein?

A

Signal transdusing kinases and cyclin dependent kinases and phosphates

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28
Q

What are the cell target proteins for EBV/EBNA1 (nuclear antigen)?

A

RBP-Jk/CBF1, glycogen synthetase

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29
Q

What are the cell target proteins for EBV/LMP1 (latent membrane protein 1)?

A

PI3K, TNF signaling components

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30
Q

What are the cell target proteins for EBV/LMP2 (latent membrane protein 2)?

A

Src family members

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31
Q

What are the cell target proteins for KSHV/k-bZIP?

A

P53

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32
Q

What are the cell target proteins for KSHV/ORF 50?

A

P53

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33
Q

What are the cell target proteins for KSHV/vCyclin?

A

Cyclin dependent kinase 6

34
Q

What are the cell target proteins for KSHV/LANA?

A

Rb family members

35
Q

What are the cell target proteins for HBV/X protein (HBx)?

36
Q

What are the target proteins for HPV/E5?

A

Epidermal growth factor

37
Q

What are the target proteins for HPV/E6?

A

P53, PD2 protein, E6-AP

38
Q

What are the target proteins for HPV/E7?

A

Rb, p21, p27, p600

39
Q

What are the target proteins for Merkel cell carcinoma/T antigen?

A

Rb, p53, Src, PP2A

40
Q

What are the characteristics of Long Latency Viruses?

A

Efficiency of formation: very low
Tumor Latency: Long
Viral Genome: Intact; replication competent
Oncogene: Virus encoded protein activates transcription
Mechanism: trans-acting protein
Transformation of Cultured cells: NO

41
Q

What is an example of Long Latency Virus?

42
Q

These are characteristics of which virus?
Genome does not contain cell oncogenes
Does not induce insertion of mutagenesis
genes are not expressed in all clones

A

HTLV associate ATLL

43
Q

T/F

Expression of viral proteins is necessary for initiation of transformation but not for its maintenance

A

False

Expression of viral genes not proteins

44
Q

What is the oncogenic protein for HTLV

A

Tax (tras acting)

Chromosomal aberration and genomic instability

45
Q

How does HTLV-1 progress after infection?

A

Infection of T cell -> cloning of CD4+ T cell by Tax -> Host genome alteration -> malignant proliferation and manifistation of ATL

46
Q

Why is the mechanism of HCV different from all other viruses?

A

It does not enter the cell nucleus. Instead it replicated in the cytoplasm.

Mechanism of tumor formation
There is a immune attack on hepatocytes (by attacks infected and non-infected cells). After repair is when cancer forms
Look at slide 61

47
Q

Chronic infection of HCV leads to?

A

Hepatocellular carcinoma (HCC)

48
Q

T/F
The mechanism of HCC is a combination of only direct mechanism, which results in chronic oxidative DNA damage that promotes the development of mutations

A

False

Direct and indirect

49
Q

What is the pathogenesis of HCV?

A

Normal liver -> Normal liver -> chronic hepatitis -> Cirrhosis -> HCC/ESLD/Death

50
Q

What is a viral vector?

A

It is a DNA molecule used a a vehicle to carry foreign material into the cell, where it can be expressed
Modified in the lab

51
Q

What does a viral vector consist of?

A

Insert (transgene-thrapuetic gene)

Sequence that serves as a backbone

52
Q

What are the 4 major types of vectors?

A

plasmids
Viral vectors
Cosmids
artificial chromosomes

53
Q

Insertion of a viral vector into a target cell is called?

A

Transformation for bacterial cells
Transfection for eukaryotic cells
transduction for insertion of a viral vector

54
Q

What is an ideal gene therapy vector?

A

Allows efficient and selective transduction of the target cells

The vector is maintained inside the cells for necessary time

Expresses genes of interest at levels necessary for achieving therapeutic effects

Safe

55
Q

to start gene therapy what do you need to know?

A
  1. Safety regulations -
  2. Administration
    Specificity of vector
    efficiency
    rout of delivery
    time of expression
    therapeutic gene-based approach
  3. Target gene
56
Q

What is Glybera?

A

AAV-based gene therapy for lipoprotein lipase deficiency (LPLD)

5 year approval bythe European Commission in 2012, but was not approved in USA.

Associated with Aden-associated virus vector

57
Q

What is Lipoprotein lipase deficiency?

A

a rare inherited disorder which can cause severe pancreatitis
Prevalence: 1-2 per million

58
Q

What are the reasons why Glybera was not sent for renewal?

A

The product was a commercial failure.
The single dose treatment costs $1.2 millions per person.
There are very limited number of patients (estimated 150-200 patients in Europe).
Only one patient was treated in 2016. Patient was given 40 injections of AAV in the muscles.

59
Q

What are the current 4 clinical gene therapy trials using AAV vectors

A

Mutation in RPE65
Cystic Fibrosis
Hemophilia B

60
Q

Mutation in RPE65 Gene Therapy Trial

A

eye protein that causes blindness - first approved gene therapy

61
Q

Cystic fibrosis Gene Therapy trial

A

aerosol delivery to the lungs, dose-dependent distribution of vector DNA in the lungs that lasted for 90 days.

62
Q

Hemophilia B Gene Therapy Trial

A
  1. Intramuscular injection. Production of Facto IX from AAV vectors was insufficient to significantly increase FIX plasma levels to alleviate disease
  2. Systemic administration. Transient FIX expression, liver toxicity, Immune response to the cased proteins and trans gene
63
Q

What is the mechanism for CART T-Cell Therapy?

A
  1. Remove blood form patient to get T cells
  2. Make CAR T cells in the lab (Insert gene from CAR into T cell -> CAR T cell will express Chimeric antigen receptor)
  3. Grow millions of CAR T cells
  4. Infuse CAR T cells into patient
    CAR T cells bind to cancer cells and kill them
64
Q

What is the receptor on the CAR T cell?

A

chimeric Antigen Receptor (CAR)

65
Q

What are the parts of the Chimeric antigen receptor?

A
  1. CD3 TCR domains - T cell activation
  2. CD28 or 4-1BB co-stimulatory domain
  3. Hinge and transmembrane domain
  4. Antibody domain recognizes CD19 B cell antigen
66
Q

AAV type 2 is mostly used for?

A

Vetor generation

67
Q

How do yo produce recombinant adeno-associated virus vector for gene therapy?

A

Co-transfection
Second plasmid contains cased protein. First plasmid is the gene that will be packed into caspid.

Rep and Cap genes are replaced with transgene Only ITRs required in cis for genome packaging

Structural (cap) and packaging (rep) genes are expressed in trans in producing cell line

Ad genes is used for helper function to produce AAV infectious particles

68
Q

What are the pros to AAV vectors for gene therapy?

A 
Non-cytotoxic and non-pathogenic without helper-virus
Transduces non-dividing cells
Absence of all viral genes in the vector
Low immunogenicity 
Long-term transduction
Broad spectrum of infection
69
Q

What are the cons for AAV vectors for gene therapy?

A

Preexisting immunity to the virus
Small packaging capacity - 4.7 kb
Difficult to produce high yield of virus (depends on plasmid transfection)
Difficult to purify

70
Q

What are the two products based on lentiviral vector were approved by FDA in 2017?

A

Kymriah

Yescarta

71
Q

What is Kymirah use for?

A

(Tisagenlecleucel, Novartis) for treatment of
B-cell precursor ALL (Pediatric) and
Relapsed/refractory diffuse large B-cell lymphoma (DLBCL)

72
Q

What is Yescarta used for?

A

Yescarta (Axicabtagene Cioleucel, Gilead Sciences Inc.) for treatment of adult B-cell lymphoma
Patient-specific product. The process of in vitro expansion of patient cells may be challenging especially in heavy pre-treated patients.

73
Q

What are the cons of Kymirah and Yescarta?

A

High failure rate of CAR T-cell manufacturing
Heterogeneity of antitumor response
Severe toxicity (cytokine storm, neurotoxicity, B-cell aplasia)
High price of vector production

74
Q

What are the approved gene therapies?

A 
Gendicin 
Glybera 
Strimuelis 
Luxturna
Adeno-associated virus
Tisagenlecleucel
Kymriah
Axicabtagene 
Cioleucel-Yescarta
75
Q

Which gene therapy is used to treat head and neck cancer

A

Gendicin

China

76
Q

Which gene therapy is used to treat Lipoprotein lipase deficiency?

A

Glybera

Europe, UniQure

77
Q

Which gene therapy is used to treat Pediatric immune disorder?

A

Strimuelis

Europe, GlaxoSmithKline

78
Q

Which gene therapy is used to treat Mutation in RPE65 eye protein that cause blindness?

A

Luxturna
Adeno-associated virus
(USA, Spark Therapeutics)

79
Q

Which gene therapy is used to treat B cell precursor ALL and Pediatric Relapsed/refractory diffuse large B cell lymphoma (DLBCL)?

A

Tisagenlecleucel
Kymriah
(USA, Novartis)

80
Q

Which gene therapy is used to treat B cell lymphoma in adults?

A

Axicabtagene Cioleucel-Yescarta

USA, Kite Pharma Inc.

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