Vaccine Associated Sarcoma

Question 1

Lawrence et al. 2012. Masitinib demonstrates anti-proliferative and pro-apoptotic actvity in primary and metastatic feline injection-site sarcoma cells

Dyssregulation of PDGFR may play a role in feline ISS

Masitinib is selective for the PDGFR signaling pathway

Results?

Drug concentation needed?

Answer 1

Inhibited cell growth and PDGFR phophorylation in 2 cell lines

Needed to be higher to inhibit growth than to modulate ligand-induced autophosphorylating of PDGFR

Question 2

Lawrence et al. 2013. Changes in g-H2AX expression in irradiated feline sarcoma cells: an indicator of ds-DNA breaks

Determine if radiation-induced damage to ISS cells could be detected using DNA DSB detection kit

Results with irradiated cells?

Answer 2

Irradiated cells showed increase in ds-DNA breaks compared to control cells

DNA dammage can be detected using mouse anti-human mAB for g-H2AX

Question 3

Rousset et al. 2013. Clinical and low-field MRI characteristics of injection site sarcoma in 19 cats

19 cats

Tumors appearance on MRI?

Minieralization?

Tumor volume?

Contrast enhancement?

Difference between excisional and incisional biopsy?

Answer 3

Hyperintense on T1W and T2W

Larger tumors more likely to have mineralization

Could not predict tumors free margins at definitive surgery

Moderate to marked contrast enhancement

Infiltrative margins and presence of T2W hyperintense zone were more prevalent following excisional biopsy while cavitation more prevalent with incisional biopsy

Question 4

Ladlow et al. 2013. Injection site-associated sarcoma in the cat: treatment recommendations and results to date

Metastatic lung involvement?

Recurrence?

Age?

Answer 4

25%

Even with clean margins tumor recurrence occurs in 1/3

Younger cats with peak presentation 6-7 years and second peak 10-11 years

Question 5

Travetti et al. 2013. CT characteristics of FSA - a histological subtype of feline ISS

Location of FSA?

Contrast enhancement?

How should the cat be positioned if tumor is intrascapular?

Answer 5

Most were interscapular (45%)

Strong, ring (42%), heterogenerous (36%), homogeneous (9%)

Per posr-contrast scan - Forelimbs positioned caudally along the body in addition to standard protocol with the forelimbs extended cranially

Question 6

Dean et al. 2013. The incidence of feline injection site sarcoma in the UK

America - 1 case per 1,000-10,000

Incidence risk per year?

Answer 6

1/16,000-50,000 cats registered by practices, 1/10,000-20,000 consultations and 1/5,000-12,500 vaccination visits

Incidence of FISS in UK is very low

Question 7

Nolan et al. 2013. Stereotactic body RT for the treatment of ISS in cats: 11 cases

Toxicity?

Later radiation toxicity?

Objective RR?

Median PFI?

MST?

Answer 7

Infrequent and limited to mid self-limiting dermatitis and colitis in 2 and 1 of 11 cats

None

8 of 11 cats (73%) - PR or CR determined by CT or PE

242 days (8 mo)

301 days (10 mo)

Question 8

Canwardine et al. 2014. UK owner preference for treatment of feline ISS

208 respondents

How many would pursue surgery? Would not?

Not allow amputations?

Would not pursue surgery for interscapular, chest, or abdomen?

Answer 8

39%, 1% would not

forelimbs (20%), hindlimb (15%), tail (15%)

26, 32, 27% would not have surgical tx of interscapular, chest, or abdomen

Willing to travel 100 miles for RT or chemo (66% and 69%)

Current recommendations may not be appropriate for UK owners

Question 9

Cantatore et al. 2014. Factors influencing would healing complications after wide excision of injections site sarcomas of the trucks of cats

Analyze factors influencing development of wound healing complication (WHC) undergoing wide excision of ISS

Main factor associated with risk of total and major WHC?

What were the significant prognostic factors for major WHC?

What influenced duration of surgery?

Answer 9

Surgical time

Pattern of reconstruction, CT dimesions, clinical dimention, weight and BCS

Excision pattern and tumor CT dimentions

Question 10

Turek et al. 2014. Masitinib mesylate does not enhance sensitivity to radiation on 3 feline ISS cell lines under normal growth conditions

Masitinib was investigated as radiosensitizer in 3 cell lines

What induced reduction of clonogenic survival?

Survival from combined masitnib and RT vs. RT alone?

Answer 10

Radiation administered alone or in combination with masitinib induced dose-dependent reduction in clonogenic survival

Survival from combined masitnib and RT was not significantly different from RT alone

Masitinib does not enhance radiosensitivity

Question 11

Hill et al. 2014. In vitro efficacy of doxorubicin and etoposide against feline ISS cell line

Cell death by single agent or combinations?

Cell cycle difference after treatment?

Answer 11

Single agent and combination drug increased cell death significantly reduced the number of viable cells

Cells in G0/G1 were reduced and G2/M were increased after treatment

Combining doxorubicin and etoposide at the lower EC yielded comparable results to the EC50 of either drug alone in degree of cytoxocity, level of apoptosis, and % of cells in G2/M phase

Question 12

Petznek et al. 2014. Murine xenograft model demonstrates significant radiosensitising effect of liposomal doxorubicin in a combination therapy for feline injection site sarcoma

Recurrence rate after surgery is 70%

Pretreatment with doxorubicin resulted in?

Results suggest?

Answer 12

Significant enhancement radiaition induced cell death - result was confirmed in in vivo

Results suggest use of cocomittant chemo-radiaiton therapy for adjuvant treatment of FISS

Question 13

Wojcik et al. 2015. Enhancing anti-tumor efficacy of Doxorubicin by non-covalent conjugation to gold nanoparticles - in vitro studies of feline FSA cell lines

Colloid gold particle are promising to overcome multidrug resistance

What was observed with Au-GSH-Dox in 4 cell lines?

What was this correlated with?

Answer 13

Higher cytotoxic effect than that of free doxorubicin in 3 out of 4 cell lines

Correlated to the acitvity of glycoprotein P

Question 14

Hartmann et al. 2015. Feline injection site sarcoma: ABCD guidelines on prevention and management

Metastatic rate?

What is a trigger for malignant transformation?

Injections of what drugs have been associated with sarcoma formation?

What is particularly linked to FISS?

Risk if lower with?

What kind of vaccines should be used?

Answer 14

10-28%

Chronic inflammatory reactions

Injections of long acting drugs (glucocorticoids)

Adjuvant vaccines that induce intense local inflammation

Modified live and recombinant vaccines

Non-adjuvanted, modified-live, recombinant vaccines

Question 15

Nemanic et al. 2016. Microscopic evaluation of peritumoral lesions of feline ISS identified by MRI and CT

All cats received CT and MRI of FISS followed by wide surgical excision

A total of 87 prei-tumoral lesions were examined

How were the lesions categorized?

How many lesions were seen on both imaging modalities at same location?

Unique imaging characteristics were seen how frequently?

Answer 15

Neoplastic (20%), non-neoplastic (59%), equivoval (22%)

25 instances

5/17 (30%) neoplastic peritumoral lesions

Extensive overlap between imaging features of neoplastic and nonneoplastic lesions preclues definitive identification of neoplastic pretumoral FISS using CT or MRI

Question 16

Bray et al. 2016. Neoadjuvant and adjuvant chemotherapy combined with anatomical resection of feline ISS: results in 21 cats

21 cats with primary or recurrent FISS received 3 cycles of neoadjuvant chemotherapy with epirubicin, then resection then another 3 cycles of chemotherapy

Median follow up time

How may developed local tumor recurrence?

MST?

Answer 16

1072 days (3 yr)

14% (3 cats) at 264, 664, and 1573 days after surgery

MST could not be calculated 80% were alive or were censored due to death from other causes

When compared to historical controls, superior rates of tumor free survival and DFI

Question 17

Terry et al. 2016. Changes in the dimension and volume of feline injection-site sarcomas following formalin fixation as determined by use of the elllipsoid volume formula and 3D- CT softwares

Shrinkage of FISS following excision and formation fixation was small and may be less than that of grossly normal tissue

Tumor volume estimated by the ellipsoid formula was consistently less than that estimated by 3D-CT software and should not be used when accuracy of tumor volume is of concern

Question 18

Porceletto et al. 2017. Feline Injection site sarcoma

Investigate IHC expression of MMP-2, MMP-9 (Gelatinases), and TIMP2 (inhibitor) as prognostic factors

Expression?

Difference b/w recurrent and non-recurrent group?

Useful as prognostic factors?

What could be evaluated as prognostic factors?

Answer 18

Variable expressed - factors play a role in tumor invasiveness

None

No

Size of tumor measure after formalin fixation with cutoff of 3.75 cm (accuracy 86%), mitotic count optimal cutoff 20/10 (accuracy 80%)

Question 19

Terry et al. 2017. Quantification of surgical margin length changes after excision of feline injection site sarcomas - a pilot study

5 cats

When did the largest mean decrease occured in gross normal surgical margins (GNSM) occur?

Formaline fixation, trimming, and mounting on slide did it affect GNSM?

Mean histologic tumor free margin was decreased when?

Answer 19

Immediately after excision

No

In vivo GNSM and on slide GNSM

Significant decrease in surgical margin occur immediately after excision (prior to fixation).

Subgross evaluation of tumor free margins from on-slide GNSM to HTFM overstimates the actucal tumor free margins.

Question 20

Ferrari et al. 2017. Clinical and CT tumor dimension assessments for planning wide excision of injection site sarcomas in cats: how strong is the agreement?

The discrepancy between clinical and CT measurement of dimension in resectable tumors has led to bias that affects surgical dose

53 cats

What did the CT measurements show compared to clinical dimensions? Increased with?

Answer 20

Tended to be larger than clinical dimensions and this increased with increasing tumor size

Question 21

Pierro et al. 2017. Anti-proliferative effect of metformin on a feline injection site sarcoma cell line independent of Mtor inhibition

Metformin is oral hypoglycemic drug that inhibits cancer cell proliferation via upregulation of AMPK (AMP activated protein kinase) and possibly inhibition of mTOR

Feline ISS cell line were treated with metformin at varied concentrations

What was observed?

Mechanism of cell death?

Was there inhibition of mTOR?

Answer 21

Dose dependent decrease in cell viability

Apoptosis or necrosis

No inhibition of mTOR or phosphorylated mTOR

Metformin causes cell death by apoptosis or necrosis but does not appear to be mediated by mTOR inhibition

Question 22

Maxwell et al. 2017. In vitro chemosensitivity of feline injection site-associated sarcoma cell lines to carboplatin

What was the effect of carboplatin on FISS cell viability?

Answer 22

Caused dose-dependent and time-dependent effects on cell viability

Local tumor control might be achived by implantation of Cl-CSH beads immediately following radical or marginal excision of the primary tumor or by implantation without tumor resection

Question 23

Longo et al. 2017. Dynamic tomographic studies of interscapular feline ISS: essential or useless practice?

Tumor volume estimates were compared between the ellipsoid and the semi-automated segmentation methods - 2 observers

How may invaded the adjacent muscular structures from 84 cats? How many muscles infiltrated?

Between the extended and flexed position the average estimated number of infiltrated muscles, was there agreement?

How were the higher tumor volumes detected?

Tumors with small volumes showed?

Answer 23

59/84 (70%), 15 muscles

Yes, 1.9 (extended) and 1.84 (flexed) and 1.89 extended and 1.85 flexed

Ellipsoid method

Slightly decreased muscle infiltrastion

Marked difference in the recorded distance between the skeletal structures and the neoplasm in the 2 different positions were established

Question 24

Muller et al. 2017. Curative intent radical en bloc resection using a minimum or a 3 cm margin in feline ISS: a retrospective analysis of 131 cases

Treated with 3 cm lateral margins (chest and abdominal wall) and 2 fascial planes (interscapular tumors)

Median DFI and ST?

Who was more likely to die?

RR and DFI for the different tumor locations?

Local tumor recurrence rate?

Who was more likely to get tumor recurrence?

Which tumor bed bodies were associated with lower RR?

Answer 24

21 and 24 months

Patients operated for recurrent tumors than de novo tumors

Comparable

38%

Recurrent tumors (RR 55.5% vs 33.3%)

Free to tumor cells were associated with lower RR compared to those with tumor cells (3.5% vs. 76%)

Question 25

Holtermann et al. 2017. The tyrosine kinase inhibitor toceranib in feline injection site sarcoma: efficacy and side effects

18 cats with unresectable FISS were treated at 3.25 mg/kg EOD

Response?

Answer 25

No clinical response :(

Question 26

Benton et al. 2017. Gold nanoparticles enahnce radiation sensitization and suppress colony formation ina feline injection site sarcoma cell line, in vitro

Assess effects of gold nanoparticles (AuNP) on ISS cytotoxicity and colony formation as standalone or combination with RT

What was the effect of AuNP of cell proliferation?

Viability and cell cycle of ISS?

Results when AuNP and RT were combined?

Answer 26

Significantly decreased cell proliferation

Not significantly altered

Decreased colony formation compared to RT alone

Question 27

Carneiro et el. 2018. Feline injection site sarcoma: Immunohistochemical characteristics

Analyzed for vimentin, cytokeratin, desmin, S100, viral leukemia virus, COS-2, c-KIT

Desmin?

Vimentin?

S-100 protein?

c-KIT?

COX-2

FeLV viral particles?

Answer 27

81% negative

100% positive

95.2% positive

19% positive

61. 9% positive
62. 9% positive

Question 28

Rossi et al. 2018. Comparison of definitive intent finely fractionated and palliative intent coarsely fractionated radiotherapy as adjuvent treatment of feline microscopic injection site sarcoma

Sx then treat with one of the RT protocol

59 cats: 38 finely fractionated protocol (48-52.8 Gy over 4 weeks) and 21 coarsely fractionated group (36 Gy over 3 weeks in 6 fractions)

PFI between the 2 groups?

Overall PFI?

When only first-occurence cases were included, median PFI?

In cats that underwent multiple surgeries, PFI?

Answer 28

Not different

2000 days

longer in finely fractionated compared with coarsely fractionated (1430 vs. 540 days)

No difference between protocol (233 vs. 395 days)

Cats with primarily occuring ISS benefit from finely fractionated RT

Question 29

Graf et al. 2018. Feline injection site sarcomas: Data from Switzerland 2009-2014

Marked decrease in incidence of FSA and occured after non-adjuvanted FeLV virus was introduced

Alum adjuvanted vaccines are involved in genesis of FISS

Question 30

Abdelmageed et al. 2018. Feline vaccine sarcomagenesis: Is there an inflammation independent role for aluminum?

Aluminum hydroxide preparation in CHO cells?

Which cells were sensitive?

Answer 30

Cytotoxic and mutagenis in CHO cells in vitro

Cells deficient in DNA ds breaks than ss break - Al is associated with ds breaks

Question 31

Hsueh et al. 2019. Role of nuclear factor-kappa B in feline injection site sarcoma

Nuclear expression of NF-κB p65 was detected in ____% of FISS cases and not correlated with tumor grading, sex, and age.

The NF-κB inhibitor, DHMEQ, was able to prevent nuclear translocation of NF-κB p65, inhibit cell proliferation, migration, and colonization in dosage-dependent manners, and induce cell apoptosis in these primary FISS cells.

Conclusions: High expression rate of nuclear NF-κB p65 in FISS cases and dose-dependent inhibitory effects on the growth of FISS primary cells treated with NF-κB inhibitor suggested that NF-κB might be a potential molecular therapeutic target for FISS.

Answer 31

83.3%

Question 32

Wei et al 2019. Elucidating the transcriptional program of feline injection-site sarcoma using a cross-species mRNA-sequencing approach

Background: Feline injection-site sarcoma (FISS), an aggressive iatrogenic subcutaneous malignancy, is challenging to manage clinically and little is known about the molecular basis of its pathogenesis. Tumor transcriptome profiling has proved valuable for gaining insights into the molecular basis of cancers and for identifying new therapeutic targets. Here, we report the first study of the FISS transcriptome and the first cross-species comparison of the FISS transcriptome with those of anatomically similar soft-tissue sarcomas in dogs and humans.

Methods: Using high-throughput short-read paired-end sequencing, we comparatively profiled FISS tumors vs. normal tissue samples as well as cultured FISS-derived cell lines vs. skin-derived fibroblasts. We analyzed the mRNA-seq data to compare cancer/normal gene expression level, identify biological processes and molecular pathways that are associated with the pathogenesis of FISS, and identify multimegabase genomic regions of potential somatic copy number alteration (SCNA) in FISS. We additionally conducted cross-species analyses to compare the transcriptome of FISS to those of soft-tissue sarcomas in dogs and humans, at the level of cancer/normal gene expression ratios.

Results: We found: (1) substantial differential expression biases in feline orthologs of human oncogenes and tumor suppressor genes suggesting conserved functions in FISS; (2) a genomic region with recurrent SCNA in human sarcomas that is syntenic to a feline genomic region of probable SCNA in FISS; and (3) significant overlap of the pattern of transcriptional alterations in FISS with the patterns of transcriptional alterations in soft-tissue sarcomas in humans and in dogs. We demonstrated that a protein, BarH-like homeobox 1 (BARX1), has increased expression in FISS cells at the protein level. We identified 11 drugs and four target proteins as potential new therapies for FISS, and validated that one of them (GSK-1059615) inhibits growth of FISS-derived cells in vitro.

Conclusions: (1) Window-based analysis of mRNA-seq data can uncover SCNAs. (2) The transcriptome of FISS-derived cells is highly consistent with that of FISS tumors. (3) FISS is highly similar to soft-tissue sarcomas in dogs and humans, at the level of gene expression. This work underscores the potential utility of comparative oncology in improving understanding and treatment of FISS.

Question 33

Fleming et al. 2019. CT angiography and MRI imaging features do not predict the tumor type and grade of feline injection site sarcoma

Computed tomographic angiography (CTA) and magnetic resonance imaging (MRI) have been described as methods for preoperative surgical planning in cats with feline injection site sarcomas (FISS), however, few published studies have compared these modalities. The objective of this retrospective, secondary analysis study was to determine if imaging features of FISS on CTA and MRI are predictive of neoplastic peritumoral projections. Archived data from a previous prospective study were retrieved for 10 cats with FISS. All cats had been evaluated in a single anesthetic episode with MRI and dual phase CT (CTA) imaging followed by surgical removal. Histopathological grading and targeted histopathology of imaging-identified peritumoral projections were performed. Two observers evaluated the CTA and MRI studies for FISS shape, margination, size, enhancement pattern, postcontrast uniformity, pre- and postcontrast margination, the number of muscles involved, mass mineralization, and bone lysis. Metal was present in the imaging field of three of 10 cats, resulting in one nondiagnostic MRI.

Peritumoral projections were detected in all cats with both imaging modalities, and most were benign. At least one neoplastic peritumoral projection was detected in six cats using MRI, five cats using CTA, and three cats with both modalities. Higher grade FISS were larger than low grade using MRI, and FISS were larger using MRI. Other FISS imaging features using MRI and CTA were similar. Findings supported use of either MRI or CTA for detecting neoplastic peritumoral projections in cats with FISS. Authors recommend CTA for cats with known metallic objects in the scan field.