Withrow and tumor types

Question 1

Fill in the blank for the increased risk with Environmental tobacco smoke:

1. Cats ___ X more likely to develop LSA
2. Cats ___ X more likely to develop OSCC, correlates with increased p53 expression
3. Dogs ___ X more likely to develop LSA or sinonasal cancer

Answer 1

1. 2.4 X
2. 2 X
3. 3.4 X

Question 2

Fill in the blank for risk factors for TCC:

1. ___ X risk of TCC in dogs exposed to herbicides alone
2. ___ X risk with exposure to both herbicides and insecticides
3. T/F: Insecticide use alone was risky?
4. Phenoxyherbicides (2,4-D, MCPA, MCCP)= ___ X risk
5. ____ x risk in dogs treated with topical insecticides, especially overweight or obese female dogs
6. Females ___ X higher risk
7. Scotties ___ X higher risk
8. veggies ___ X lower risk

Answer 2

1. 3.6
2. 7.2
3. FALSE
4. 4.4
5. 28
6. 27
7. 18-21
8. 0.3

Question 3

Fill in the blank for risk factors for Rural vs. urban environment:
1. Urban pets= increased risk of what K9 cancers?

2. ___ X increased risk of canine LSA in cities with illegal waste dumping
3. Newer study found increased risk of what cancer with proximity to waste incinerators, polluted sites, radioactive waste?

Answer 3

1. LSA, tonsillar SCC, nasal carcinoma
2. 2.4 X
3. LSA

Question 4

Fill in the blank for risk factors for hormonal factors:

o Spay/neuter and mammary cancer

1. Dogs: ___ % risk if before 1st estrus, ____% if after 1st estrus, ___ if after 2nd heat
2. Cats: ___ % reduced risk if spayed < 6 mo (7X lower risk), ___ % reduction if spayed < 1 yr
3. Intact or Neutered female dogs less likely to develop LSA? (choose one)
4. Intact or neutered males have 1.4 X increased risk of AGASACA? (choose one)

o Prostate cancer:

5. Neutered dogs have ___ x increased risk
6. T/F: Risk association highest for prostatic TCC
7. Urinary TCC: ___ X increased risk with neutering
8. Dog breeds associated with risk of neutering and urogenital tumors?

Answer 4

1. 0.05%, 8%, 26%
2. 91%, 86%
3. Intact
4. Neutered
5. 2.8-3.4
6. True
7. 3.6 X
8. mixed breed, Dobie, Sheltie, Scottie, beagle, GSP, Airedale, Norwegian elkhound

Question 5

Fill in the blank for risk factors for chronic inflammation:

1. Dogs with anthracosis (black dust) in their lungs = ___X increased risk of primary lung tumors
2. Dogs with hx of TPLO were ___ X more likely to develop proximal tibial OSA

Answer 5

1. 2X

2. 40X

Question 6

Fill in the blank for risk factors cats:

• UV radiation
1. Light-colored cats exposed to UV radiation = ___ X increase risk of developing SCC

•Flea collars

2. ___ X increased risk of feline OSCC with use of flea collars
3. Increased risk with high intake of either canned food= ___ X or canned tuna fish= ___ X

Answer 6

1. 13.4
2. 5 X
3. 3 X, 4.7 X

Question 7

Fill in the blank for risk factors for obesity:

1. Increased risk of which cancers?
2. CMTs= obesity during which life stage increases risk?
3. TCC= T/F: Obesity further increases risk in dogs exposed to topical insecticides?

Answer 7

1. canine mammary tumors and TCC
2. puberty
   overweight dogs more likely to have lymphatic invasion and grade III tumors
   Estrogen is produce by adipose tissue
3. True

Question 8

What is Agnor and where does it stain?

Answer 8

AgNor = argyrophilic nucleolar organizer region proteins
**nucleus at the centrosome
Nuclear proteins that accumulate in highly proliferating cells

Question 9

1. What is a gene gun?
2. What is suicide gene therapy?
3. What is it limited by?

Answer 9

1. Gene gun is a means of transfection (introduction of DNA plasmid into cells); in this case, DNA is coupled to a nanoparticle of an inert solid and “shot” directly into the target cell. Typically DNA is attached to gold particle, which independently penetrates cell membrane. Once in the cell, DNA dissociates from the particle and can be expressed.
2. Suicide gene therapy (aka gene-directed enzyme/prodrug therapy [GDEPT]) = a gene encoding for a non-mammalian enzyme is able to convert a nontoxic prodrug into a toxic metabolite, allowing for localized delivery to cancer cells. As a result, the activated drug demonstrates cytotoxic effects on cancer cells as well as non-transfected cancer cells (bystander effect).
3. Limited by degree of tissue penetration

Question 10

What are the advantages of gene therapy with a gene gun?

Answer 10

(1) freedom from complex biologic systems (i.e., viruses) and toxic chemicals (i.e., liposomes)
(2) delivery is independent of cell membrane receptors (3) allows delivery of various sizes of DNA, incl. large ones
(4) lacks extraneous DNA or proteins
(5) possibility for repetitive treatments

Question 11

What was the outcome for suicide gene delivery in K9 melanoma when dogs were injected with irradiated xenogeneic cells with human IL-2 and GM-CSF post-op?

Answer 11

Improved % disease-free and % met free. When used in the gross disease setting, 49% ORR

Question 12

Vaccine-Associated Feline Sarcoma Task Force: What were the guideline recommendations with vaccination?

Answer 12

When to biopsy:

(1) Mass increases in size for >1 month post vaccine
(2) Mass is >2 cm in diameter
(3) Mass has been present for 3 months after

Question 13

Vaccine-Associated Feline Sarcoma Task Force: What were the temporal changes noted with change in vaccination?

Answer 13

Temporal changes:
1. DECREASE – interscapular, lateral thoracic wall, and tumors cranial to the diaphragm
 Interscapular tumors still the majority of all ISS (40%)
2. INCREASE – right forelimb (NOT left), right pelvic limb, left pelvic limb, abdominal wall, and tumors caudal to the diaphragm
o Patient signalment and tumor histotype did NOT change
o Assuming causality based on location, rabies vaccine accounts for 52% and FeLV 29%

Question 14

1. What is the prognosis or MST for oral SCC in dogs with sx, RT, and/or chemo?
2. What about with no tx?
3. In another paper that found sx alone did not statistically improve survival, what was the MST for sx and chemo, and then for sx, chemo, and RT?
4. What are 2 negative px indicators for oral SCC?
5. What % of dogs and cats are considered to have systemic dz at the time of dx with oral SCC?
6. What % of dogs with oral SCC can gain local control with RT alone, but only about 10% will be alive at 1 yr?

Answer 14

1. MST 179 days (6 mo) regardless of treatment
2. No treatment: MST 65 days
3. Surgery + chemo: MST 212 days (7 mo)
   Surgery + RT + chemo = MST 355 days (12 mo)
4. Anorexia and lethargy were negative prognostic indicators
5. Considered systemic in 90% of dogs and cats at diagnosis (10-20% pulmonary mets)
6. RT: 75% local control but 10% 1-year survival

Question 15

1. What is the most common benign hepatobiliary tumor in the cat?
2. What is the most common malignant tumor?
3. What is the metastatic rate of a bile duct carcinoma in a cat?
4. Hepatobiliary tumors in cats may be caused by what?
5. In cats, majority of hepatocellular tumors are benign or malignant?
6. T/F: Azotemia is very common in cats with liver tumors and may occur in the absence of liver enzyme elevation.

Answer 15

1. Biliary cystadenomas (52%)
2. bile duct carcinoma
3. 67-80% metastatic rate in cats
   (side note: 2nd most common malignant tumor in dogs- 88% met rate)
4. trematode infections
5. adenomas (benign), rather than HCC
6. True

Question 16

What are the negative px indicators for AGASACA?

Answer 16

1. primary tumor size (> 10 cm2 or >2.5 cm LD)
2. presence of LN metastasis
3. presence of distant metastasis
4. advanced clinical stage
5. lack of surgery
6. treatment with chemotherapy alone
7. lack of treatment

Question 17

Give examples of an autocrine feedback loop in regards to co-expression of ligands and receptors in certain canine cancers.

Answer 17

1. Co-expression of MET and HGF in canine OSA

2. Co-expression of c-Kit and SCF in canine HSA

Question 18

In OSA cell lines, was normal vs. high ALP assoc. with differences in cell proliferation, migration, invasion, or chemosensitivity?

Answer 18

NO

*No difference in gene expression patterns either

Question 19

Endothelin-1 and endothelin A receptors are expressed in canine OSA cells. What is the main MOA of these receptors?

Answer 19

mediate OSA migration, survival, proliferation, and increase expression of bALP

Question 20

What has been noted about the Wnt pathway and B-catenin in OSA cells in dogs?

Answer 20

B-catenin transcriptional activity in downregulated in OSA cells, suggesting the canonical Wnt pathway is minimally activated. (Piskun et al, VCO, 2016)

Question 21

What occurs to canine OSA cells if there is decreased B-catenin transcription in regards to txt with doxo? What about with carbo?

Answer 21

Decreased B-catenin transcription = reduction in proliferation and decreased sensitivity to doxorubicin (did not affect carbo)

Question 22

Is there a difference in B-catenin signaling or localization between dogs with high vs. normal serum ALP?

Answer 22

ALP thought to be downstream of B-catenin (activated by Tcf), but no difference in B-catenin signaling or localization between dogs with high vs. normal serum ALP

Question 23

1. B-catenin is expressed in _____ % of primary OSA tumors. Wnt5a is expressed in ____ % of primary tumors.
2. Where within the cell has this expression been noted to occur?
3. Have mutations been noted, and does this affect DFI and MST?

Answer 23

1. 81%, 54%
2. Increased cytoplasmic expression (if active, B-catenin should be nuclear)
3. No mutations identified, and no effect on DFI or MST.
   Wnt5a is expressed in 54% of primary tumors, but is not assoc. with DFI or MST.

Question 24

Fill in the blank for OSA cell lines and chemokine expression:

1. Cancer cells bearing which chemokine metastasize to lung, bone, LN, and liver (sites of high CXCL12 concentrations)?
2. Canine OSA cells express which chemokine mRNA and protein; upregulated in the majority of primary tumors?
3. Which bisphosphonate decreases serum CXCR4?

Answer 24

1. Cancer cells bearing CXCR4 metastasize to lung, bone, LN, and liver (sites of high CXCL12 concentrations)
2. Canine OSA cells express CXCR4 mRNA and protein; upregulated in the majority of primary tumors
3. Zoledronate

Question 25

1. What is the MST for rib OSA with sx alone vs sx and chemotherapy? 2. What is the recommended sx proceduce for rib OSA? 3. Is increased ALP assoc with decreased survival?

Answer 25

1. MST 3 months with Sx alone; 8-9.6 months with adjuvant chemotherapy 2. Remove 1 additional rib in either direction + ~3cm dorsal and ventral margins 3. Increased ALP assoc. with decreased survival (7 months vs. 23 months)

Question 26

What is the MST for dogs with rib CSA that have sx and does this px depend on grade?

Answer 26

MST >3 years with surgery; | unlike appendicular CSA, does NOT depend on grade

Question 27

1. What is the BRAF mutation and where does it occur within the genome in the dog? 2. What is the corresponding human mutation? 3. Mutations in B-RAF leading to constitutive activation of which pathway?

Answer 27

1. T to A transversion along exon 15, resulting in AA substitution of glutamic acid for valine at codon 450 (V450E aka V595E) on chromosome 16 2. Corresponds to V600E mutation on chromosome 7 in humans 3. Mutations lead to constitutive activation of MAPK pathway

Question 28

1. What is the frequency of BRAF mutation in dogs with TCC ? 2. What is the sensitivity of BRAF detection with Sanger test? 3. What is the sensitivity of BRAF detection with ddPCR test? 4. What is the specificity of both tests?

Answer 28

1. BRAF mutation present in 87.9% of TCC overall (histopath and urine) urine detected mutation in some histopath samples that were negative 2. Sanger sensitivity o Tissue: 71% overall (67% of UC and 78% of PC) o Urine: 61% of UC and 100% of PC 3. ddPCR sensitivity o Tissue: 79% overall (75% of UC and 85% of PC) o Urine: 85% overall (83% of UC and 100% of PC) Droplet digital PCR (ddPCR) has a higher sensitivity than Sanger 4. both techniques are 100% specific o Able to detect as little as 1 in 10,000 alleles (0.01%).

Question 29

What TSG has been detected in humans and dogs with high grade LSA leading to shorter ORR and MST?

Answer 29

1. P53 mutation in humans and 16-22% of dogs with high grade LSA (shorter ORR and MST in dogs) Side note: Hypermethylation of DAPK CpG islands (a serine/threonine kinase involved in IFN-y induced apoptosis) = shorter PFS and MST in dogs and humans

Question 30

What chromosomal abnormalities have been detected in dogs with LSA?

Answer 30

1. Gain of CFA 13 and 31, loss of CFA 11 and 14 2. Trisomy of 13 = INCREASED 1st remission duration and MST 3. Loss of CFA 11 assoc. with deletion of INK4 TSG locus (incl. p16); only detected in high grade T-cell LSA

Question 31

Most common lymphoma phenotype in Boxers. | What is the typical flow cytometry in Boxers?

Answer 31

85% of LSA in Boxers is TCL | predominantly TCR αβ+, CD4+, and lymphoblastic high-grade morphology

Question 32

Which of the following is true regarding the CD40-activated B cell cancer vaccine in dogs with non-Hodgkin's lymphoma? a. Vaccination with RNA loaded CD40-B is not well-tolerated due to significant alterations in complete blood chemistries and long-term complications of vaccination such as autoimmunity b. CD40-activated B cell cancer vaccine improves second clinical remission and survival time in these patients c. RNA-loaded CD40-B cells induce functional, antigen-specific T cells from healthy dogs, but not dogs with lymphoma d. There is no evidence to show that this vaccine improves remission or survival rates

Answer 32

b. CD40-activated B cell cancer vaccine improves second clinical remission and survival time in these patients o RNA-loaded CD40-activated B-cell vaccine administered to dogs in remission after induction chemotherapy. Did not affect ORR, TTP, MST. o Dogs receiving vaccine had a higher response to rescue chemotherapy (40% vs. 8%) and longer LSS (from rescue to death)

Question 33

1. What would be the expected signalment, blood work, and FC profile of a bulldog that developed CLL? 2. What immunophenotype would be the most common?

Answer 33

1. younger (6 yr) higher % hyperglobulinemia, relative to other B-CLL lower CD25, lower MHC class II 2. May arise from a naïve, unactive B-cell

Question 34

2. A. What is the reported median survival time for dogs, independently from therapy, with CLL that develop Richter’s syndrome? a. 22 days b. 40 days c. 75 days d. 90 days B. Which of the following are common clinical signs noted in dogs diagnosed with Richter’s syndrome? i. Neurological signs ii. Lymphadenopathy iii. Diarrhea iv. Vomiting v. Weight gain a. i, ii, iv b. iii, v c. iii, iv, v d. i, iii, iv

Answer 34

Answer: A. b, B. a * Affects 5.2% of dogs with CLL; B-cell also appears most common * Lymphocyte count previously elevated with CLLs; decreased at time of RS diagnosis * Double population of small lymphs and large blasts * Concurrent cytopenias common * Rapid decrease in lymph count and onset of anemia in dogs with CLL may indicate aggressive transformation * MST 41 days, poorly responsive to CHOP

Question 35

Mechanism of hypercalcemia in multiple myeloma: IL-1beta or TGF-beta?

Answer 35

Production of osteoclast-activating factor (aka IL-1beta) by neoplastic cells Other causes: PTHrp production, TNFa, IL-6 • NOT TGF-B

Question 36

What are the etiologic factors associated with dermal HSA? 1. predisposed breeds 2. location 3. main etiology behind development

Answer 36

1. Light pigmented dogs with thin coats: Whippets, Salukis, bloodhounds, beagles, white bulldogs, English pointers 2. Typically occur on ventral abdomen 3. Histopathology = solar dermatosis consistent with UV radiation damage= actinic changes!

Question 37

Dermal HSA in dogs: 1. Risk factors for loco-regional recurrence? 2. Risk factors for metastasis? 3. T/F: Predisposed breeds less likely to develop metastasis? 4. ****Positive prognostic indicators?**** 5. Negative prognostic indicators?

Answer 37

1. predisposed breed (2.5X), ventral location, multiple masses at presentation 2. SQ involvement (2X) 3. True 4. ******Positive prognostic indicators****** o Predisposed breeds: MST 1570 days (vs. 593 days) o Ventral abdomen location: MST 1549 days (vs. 545 days) o Presence of solar changes: MST 1549 days (vs. 545 days) 5. Negative prognostic indicators o Development of metastasis or hemoabdomen: decreased MST (~540 days)

Question 38

What effect does Yunnan Baiyao have on canine hemangiosarcoma cell lines in vitro? a. Caspase-3/7 activity decreases in correlation with the IC50 in each cell line b. Yunnan Baiyao causes dose and time dependent HSA cell death through initiation of caspase-mediated apoptosis c. There is no evidence to show that Yunnan Baiyao has any effect on hemangiosarcoma cell lines d. Yunnan Baiyao increased cell grow through the NFKB pathway

Answer 38

b. Yunnan Baiyao causes dose and time dependent HSA cell death through initiation of caspase-mediated apoptosis

Question 39

How do you diagnose canine HSA without echocardiogram and only blood/urine tests?

Answer 39

Dogs with cardiac HSA had significantly higher cTnI than dogs with idiopathic pericardial effusion, HSA at other sites, or other cardiac tumors Plasma cTnI > 0.25 ng/mL is more likely cardiac HSA (sensitivity and specificity increased in pericardial effusion was present)

Question 40

1. What is the most common TSG mutation in HS? 2. What other alteration have been detected in HS? 3. FCR were ___ X more likely as BMD to have abberant RB1 and TP53. 4. A gain of function mutation in exon 3 of what gene was found in BMD?

Answer 40

1. 63% of BMD and FCR had deletions in CDKN2A/B on CFA 11 (Hedan et al, BMC Cancer, 2011). * Other alterations included deletion in RB1 (56%), deletion in PTEN (40%), and gain of function in TP53 (27%) * FCR were 2X as likely as BMD to have aberrant RB1 and TP53. * 2018: gain of function mutation in exon 3 of PTPN11 gene (substitution at the SH2 domain of SHP2) in BMD