Osteosarcoma Flashcards
Bracha et al. The expression and role of seratonin receptor 5HTR2A in canine osteoblats and an OSA cell line
5HTR2A was _____ in malignant cell line compared to normal cells
In CnOb cells, ERK-P was _____ in response to both seratonin and 5HTR2A antagonist, ritanserin
In COS cells, ERK-P ____ with both treatments
CREB was ____ in CnOb cells and ____ in COS cells
Seratonin treatment promoted ____ in malignant cells but not normal osteeoblasts
Ritanserin ____ cell viability in normal and OSA cells
Ritanserin induced ____
Oveexpressed
Decreased
Increased
undetectable, constitutively
cell viability
Reduced
Apoptosis in COS
Dailey et al. HES1, a target of Notch signaling, is elevated in canine osteosarcoma, but reduced in the most aggressive tumors
Hairy and enhancer of split 1 (HES1), a transcriptional repressor, is downstream target of Notch signaling
Notch signaling and HES1 have been linked to growth and survival
HES1 mRNA was ___ in tumor samples relative to normal bone, but ___ in tumor samples from dogs with DFI<100 days to those DFI>300d
NOTCH2 and HEY1 mRNA was ___ in tumors relative to normal bone but not differntially expressed between DFI tumor groups
Survival analysis?
Notch signaling occurs and may contribute to canine OSA. Mechanisms that do not alter HES1 expression may drive aggressive tumor
Increased
Increased, decreased
Increased
Decreased HES1 and shorter DFI
Sternberg et al. Association between absolute tumor burden and serum bone-specific alkaline phosphatase in canine appendicular OSA
96 dogs with appendicular OSA
Quantity of membranous BALP was propotional with?
In dogs devoid of macroscopic metastases there was a positive correlation between serum BALP and ___
Dog with progressive metastases, serum BALP activity ___ and coincided with development of _____
Tumor burden is determinant of serum BALP activity
Cell densitiy, not tumorigenic or metastatic phenotype
Primary tumor size
Increased, macroscopic metastases
Skorupski et al. Carboplatin versus alternating carboplatin and doxorubicin for the adjuvant treatment of canine appendiculay OSA: a randomized, phase III trial
6 doses of carboplatin or 3 doses of carboplatin and doxorubicin in alternating schedule
DFI for carboplatin vs. carboplatin and doxorubicin?
425 (14 mo) versus 135 days (4.5 mo)
Sivacolundhu et al. 2013. Ulnar OSA in dogs: 30 cases (1992-2008)
11 dogs treated with partial ulnar ostectomy and 14 with amputation, 5 dogs no resection
Median DFI and ST
Negative prognostic factors on univariable analysis?
Negative prognostic factor on multivariate analysis? MST?
437 (14.6 mo) vs. 463 days (15.4 mo)
Histologic subtype and lung metastasis
Telangiectatic or telengiectatic-mixed subtype, 208 days
Amsellem et al. 2014. Appendicular OSA in small breed dogs: 51 cases (1986-2011)
Dogs <15kg
9 treated nonsurgically, 16 amputation, 26 curative intent treatment, MST?
MST did not differ significantly between amputation and curative intent group - chemo efficacy?
For dogs in nonsurgical group, MST ____ significantly as tumor ___ increased
For dogs in amputation group, MST ___ as body weight increased
Tumor histologic score and mitotic index were lower and MST following amputation
112 days (3.7 mo), 257 days (8.6), 415 days (13.8 mo)
Decreased, tumor histologic score
Decreased
Culp et al. 2014. Evaluation of outcome and prognostic factors for dogs living greater than one year after diagnosis of osteosarcoma: 90 cases
Median age
Median weight
Serum ALP activity was high in?
Most common tumor location?
Metastatic disease?
MST beyond 1 year?
Which dogs had significantly improved survival?
8.2 years
38 kg
48%
distal portion of radius 60%
54%
243 days
Surgical site infection after limb sparing surgery
Batchinski et al. Evaluation of ifosfamide salvage therapy for metastatic canine OSA
19 dogs and 17 evaluated for response
Ifosfamide doses 375-425 mg/m2 with median of 2 doses
Overall response to ifosfamide?
Toxicity?
Median survival duration from first dose to death?
11.8%
2 dogs were hospitalized
95 days
Alvarez et al. 2014. Postoperative adjuvant combination therapy with doxorubicin and noncytotoxic suramin in dogs with appendicular OSA
Suramin increased tumor sesitivity to chemotherapy
47 dogs received 6.75 mg/kg suramin IV followed by 30 mg/m2 doxorubicin 4 hrs later - repeated q2wk x 5 doses
Median DFI
MST
203 days (6.8 mo)
359 days (1 yr)
Coyle et al. Biologic behaivour of canine mandibular OSA. A retrospective study of 50 cases (1999-2007)
How many dogs developed metastatic disease?
Median MFI
MST
In univariate analysis what was prognostic?
Grade also influence ST, ___ dogs with grade II/III tumors met free at 1 year vs. ___ for grade I
___ alive with grade II/III vs. ____ alive with grade I
Multivariable analysis, histologic grade and adjuvant chemotherapy were prognostic for MFI and MST
58%
627 days (20.9 mo)
525 days (17.5)
MI >40 decreased MFI and ST
24% vs. 72%
24% vs, 77%
Bracha et al. Evaluation of toxicities from combined metronomic and MTD chemotherapy in dogs with OSA
Tolerability of piroxicam and cyclophosphamide combined with carboplatin or caboplatin and doxorubicin MTD
All dogs underwent amputation and chemotherapy
14 dogs treated with carbo and 16 treated with carbo and doxo
Toxicities?
DFI
MST
Grade 3 and 4 in carbo group
Not significantly diffrernt 192 d in cabro and metronomic vs. 182 day
217 (7 mo) vs. 189 days (6.3 mo)
Selmic et al. 2014. Comparison of carboplatin and doxorubicin-based chemotherapy protocols in 470 dogs after amputation for treatment of appendicular OSA
carbo for 4 or 6 cycles (CARBO6), dox q14d or 21d for 5 cycles, alternating carbo and dox q21d for 3 cycles
Median DFI
MST
A lower proportion of dogs prescribed ___ experienced AEs compared to other protocols (___%)
____ was not associated with development of metastasis or death
291 days (10 mo)
284 days (9.5 mo)
CARBO6, 48%
DI - dose intensity
Duffy et al. Outcome following treatment of soft tissue and visceral extraskeletal OSA in 33 dogs: 2008-2013
Outcome after surgical treatment of non-mammary and non-thyoridal sofy tissue and visceral EOS in dogs
Most common primary tumor site?
Which dogs had longer survival times? MST?
Spleen
Wide or radical tumor excision had longer survival than marginal tumor excision - 90 days vs. 13 days
Viall et al. Antagonism of seratonin receptor 1B decreased viability and promotes apoptosis in the COS canine OSA cell line
Seratonin receptor 1B (5HTR1B) exhibits anti-proliferative activity in osteoblasts
Equal levels of 5HTR1B gene and protein expression in normal and malignant osteoblasts
Treatment with seratonin enhanced viability OSA cells but not normal osteoblasts
Anpirtoline, 5HTR1B antagonist, caused no change in viability
Specific receptor antagonist SB224289 caused reduction in osteoblast viability, effect more in OSA cells
Aberrant signaling in COS - evaluating ERK and CREB
Holmes et al. Canine OSA cell lines from patients with differing serum ALP concentration display no behaivoural differences in vitro
Difference in cell proliferation, migration, and cell invasion between cell lines with normal or increased ALP activity
No differnce was noted
McCleese et al. Met interacts with EGFR and Ron in canine OSA
Met, RTK, is overexpressed in canine OSA
In humans, RTK Met, EGFR, and Ron are coexpressed and engage in heterodimerization
Expression of EGFR and Ron in canine OSA cell lines?
TGF-a and HGF induced amplification of ERK1/2 and STAT3 phosphorylation in OSA cells and Met was phosphorylated following TGF-a stimulation - receptor cross talk
Treatment of OSA cells with gefitinib and crizotinib inhibited cell proliferation
Met, EGFR, Ron interact in OSA cells
Expressed in OSA cell lines and primary tissue, EGFR and Ron are phosphorylated in OSA tumor samples, and Met is co-associated with EGFR and Ron in cell lines
Maniscalco et al. PDGF and PDGFRs in canine OSA: new targets for innovative therapeutic startegies in comparitive oncology
PDGF and PDGFRs overexpressed in 70-80% human OSA
33 canine samples
Expression of PDGF-A and PDGF-B in OSA?
Expression of PDGFR-A and PDGFR-B in OSA?
qPCR showed all canine OSA cells overexpressed PDGFR-a while 6/7 overexpressed PDGFR-b and PDGF-A compared to normal osteoblastic cell line
PDGFR inhibitor casue dose- and time-dependent decrease in AKT phosphorylation - PDGFRs/PDGFs are co-expressed suggest autocrine/paracrine loop
40% and 60%
78% and 81%
Bergeron et al. The effect of Zhangfei on the unfolded protein response and growth of cells derived from canine and human OSAs
Protein zhangfei could suppress unfolded protein response (UPR) and growth of OSA cells
Zhangfei expressing D-17 cells displayed large vacuoles and expressed phosphatidylserine on external surface - induced micropinocytosis and apoptosis
Zhagfei inhibited thapsigargin-induced UPC in D17 cells
Ability of Zhangfei to suppress UPR and tumor cell growth may not be linked
Loftus et al. The 5-lipooxygenase inhibitor tepoxalin induced oxidative damage and altered PTEN status prior to apoptosis in canine OSA cell lines
Tepoxalin treated cells undergo apoptosis through ____ activation
Apoptosis is superceded by _____
5-LOX inhibition also increased ____ activity by preventing its alkylation or oxidation
PTEN modification allows PIP3 heigtening AKT phosphorylation
Target oxidation and LOX inhibition play roles in apoptotic response
Caspace-3
ROS
PTEN
Piskun et al. B-catenin transcriptional activity is minimal in canine OSA and its targeted inhibition results in minimal changes to cell line behaivour
Impact of inhibiting canonical Wnt signaling in canine OSA - B-catenine siRNA or dominant negative T-cell factor construct
There were no changes to cell lines with either construct
B-catenin activity was ___ in normal canine osteoblasts compared to canine OSA cell lines
Wnt signaling has minimal activity in canine OSA and targeted inhibition is not relevant
3-fold higher
Portela et al. Pro-tumorigenic effects of transforming growth factor beta 1 in canine OSA
TGFB1 contributes to skeletal remodeling inducing osteoblast proliferation, migration, and angiogeneis
33 dogs with appendicular OSA
Canine OSA cells secrete ___, express ___, and TGFB1 signaling blockade _____
Naturally occuring OS samples abundantly express ______
OS-bearing dogs, circulating TGFB1 concentration correlated with ___
TGFB1 inhibitors may impede OS progression
TGF1B, cognate receptors, TGF1B, decreased proliferation, migration, and VEGF secretion
TGFBR1/II
N-telopeptide excretion
Shoeneman et al. Survivin inhibition via EZN-3042 in canine lymphoma and OSA
Survivin inhibits apoptosis and drives cell proliferation - elevated in human and canine cancer
Survivin inhibition with EZN-3042 inhibits growth, induced apoptosis, enhances chemosensitivity in vitro
Survivin directed therapies may be effective in treatment of canine LSA and OS
Szigetvari et al. Wnt5a expression in canine OSA
Wnt5a protein, ligant for non-canonical Wnt, implicated in aggressiveness of cancer cell lines, including those from human OSA origin
Wnt5a expression in canine OSA?
Association with PFS?
54%
Wnt5a was not associayed with PFS
Neumann et al. 2015. The association of endothelin-1 signaling with bone alkaline phosphatase expression and protumorigenic activities in canine OSA
Endothelin-1 is active during bone repair, and responsible for osteoblast migration, survival, proliferation, and bone alkaline phosphatase expression
45 dogs with appendicular OSA
Expression of endothelin-1 and endothelin-A receptor?
What does the signaling mediate?
Circulating bone ALP activities were positively correlated with primary tumor bone mineral densities
Expressed in canine OS cells
Survival, proliferation, and bone alkaline phosphatase activities
Rodrigues et al. OSA tissues and cell lines from patients with differing serum ALP concentration display minimal difference in gene expression patterns
Microarray analysis in 18 OSA samples and 6 OSA cell lines from dogs with normal and increased ALP
No difference in gene expression patterns were noted.
ALP concentration is not associated with intrinsic differences of OSA cells
Oblak et al. The impact of pamidronate and chemotherapy on survival times with appendiculat primary bone tumors treated with palliative RT
RT alone, RT + chemotherapy, RT + pamidronate, and RT + chemotherapy + pamidronate
MST was longest for which dogs?
MST shortest for which dogs?
Difference in MST for dogs who receieved ___ and those that did not was statistically signifcant in univariate analysis
The addition of ___ into any protocol improved survival?
RT and chemotherapy - 307 days (10 mo)
RT and pamidronate - 69 days
Pamidronate
Chemotherapy
Helmerick et al. The effects of baicalein on canine osteosarcoma cell proliferation and death
Baicelein is a flavanoid with anti-inflammatory and antineoplastic properties
Canine OSA cell lines were treated with Baicelein
Modest ____ changes were observed in 1 cell line
Effective in inducing ___ and did not prevent doxorubicin cell proliferation inhibition
Cell cycle
Apoptosis
Thayanithy et al. 2012. Combinatorial treatment of DNA and chromatin-modifying drugs cause cell death in human and canine osteosarcoma cell lines
Downregulation of microRNAs (miRNAs) at the 14q32 locus stabilizes the expression of cMYC, thus significantly contributing to osteosarcoma (OS) pathobiology. Here, we show that downregulation of 14q32 miRNAs is epigenetically regulated. The predicted promoter regions of miRNA clusters at 14q32 locus showed no recurrent patterns of differential methylation, but Saos2 cells showed elevated histone deacetylase (HDAC) activity. Treatment with 4-phenylbutyrate increased acetylation of histones associated with 14q32 miRNAs, but interestingly, robust restoration of 14q32 miRNA expression, attenuation of cMYC expression, and induction of apoptosis required concomitant treatment with 5-Azacytidine, an inhibitor of DNA methylation. These events were associated with genome-wide gene expression changes including induction of pro-apoptotic genes and downregulation of cell cycle genes. Comparable effects were achieved in human and canine OS cells using the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA/Vorinostat) and the DNA methylation inhibitor Zebularine (Zeb), with significantly more pronounced cytotoxicity in cells whose molecular phenotypes were indicative of aggressive biological behavior. These results suggested that the combination of these chromatin-modifying drugs may be a useful adjuvant in the treatment of rapidly progressive OS.
won Seo et al. Fluoroquinolone-mediated inhibition of cell growth, S-G2/M cell cycle arrest, and apoptosis in canine OSA cell lines
Most patients are treated with flouriquinolone who develop post-operative infections
Ciprofloxacin or enrofloxacin ____ expression leading to ____?
Flouroquinolone exposure induced ____ of canine OSA cells as demonstrated by ______ and ____, and activation of _____
Inhibited p21WAF1, decreased proliferation and increased S-G2/M accumulation
Apoptosis, cleavage of caspace-3, PARP, caspace 3/7
Simcock et al. 2012. Evaluation of a single subcutaneous infusion of carboplatin as adjuvant chemotherapy for dogs with OSA: 17 cases
Limb amputation or limb-sparing surgery followed by single SC infusion of carboplatin (total dose 300 mg/m2 over 3, 5, 7 days)
MST?
AEs?
No significant difference was found in ST of dogs grouped to tumor location, type of surgery, duration of carboplatin infusion, or postoperative infection
365 days
9 had hematological effects, 1 GI, 7 infections at surgical site
Maeda et al. Genomic instability and telomere fusion of canine OSA cells
All 8 cell lines increased abnormal metacentric chromosome and telomere fusion and interstitial telomeric signals
Evidence of unstable telomeres, colocalization of g-H2AX and telomere signals in interphase cells
All canine OSA samples were telomerase positive
Telomere fusion may serve as potential marker for canine OSA
Bongiovanni et al. IHC investigation of cell cycle apoptosis regulators (Survivin, B-catenin, p53, caspace 3) in canine appendicular OSA
B-catenin staining?
Nuclear survivin and p53?
Moderate/high B-catenin expression was significantly associated with ______?
Moderate/high nuclear p53 expression was associated with ___?
Moderate/high nuclear survivin showed tendency toward ___?
p53 was negative prognostic marker and survivin potential positive prognostic indicator (confusing)
Nuclear staining detected in normal osteoblasts adjacent to tumor in 47% of cases. Cytoplasmic and membranous staining was also observed
Positive in all cases
Development of metastasis
Moderate/high histologic grade and shorter OS
Longer OS
Ryseff et al. Detection of ALP in canine cells previously stained with Wright-Giemsa and its utility in differentiating OSA from other mesenchymal tumor
83 dogs, 17 OSA and 66 other tumors (amelonotic melanoma, GIST, collision tumor, and anaplastic sarcoma)
ALP expression?
Sensitivity and specificity?
15/17 OSA and 4/66 other tumors expressed ALP
88% and 94%
Boerman et al. Prognostic factors in canine appendicular OSA - a meta-analysis
What were negative prognostic factors?
What was associated with shorter ST and DFI?
Elevated ALP and proximal humerus for both ST and DFI in dogs with appendicular OSA
Increasing age, however was not statistically significant
Selvarajah et al. Expression of EGFR in canine OSA: association with clinicopathological parameters and prognosis
EGFR mRNA and protein expression in OS cell lines, normal bones, frozen primary OS and tissue microarrays
EGFR expression in primary OS?
Tissue microarray analysis revealed thay subset of canine OS with high EGFR was associated with ___ ?
Cytoplasmic expression of EGFR?
Subset of dogs may benefit from anti-EGFR therapies
Significantly elevated in OS compared to normal bones and in OS metastasis to the lung in comparison to extrapulmonary sites
Significantly shoter survival times and DFI
75% of metastasis and similar in primary tumors
Karnik et al. 2012. Accuracy of CT in determining lesion size in canine appendicular OSA
Statistically significant association with good correlation between true lenght of OSA compared to lenght of intramedullary/endosteal abnormalities on CT
Not significant association between tumor lenght and lenght of periosteal proliferation
Statistical significant association but poor correlation between tumor lenght compared to lenght of abnormal contrast enhancement
Wasserman et al. Suppression of canine myeloid cells by soluble factors from cultured canine tumor cells
Canine BM-DCs and DH82 cell line shared?
Myeloid cells exposed to tumor derived soluble factors (TDSF) showed?
Soluble factors produced by tumor cells suppress activation and function of myeloid cells.
CD11b, CD11c, and MHCII expression
Decreased MHCII expression and CD80 - reduced phagocytic activity and suppressed proliferation of responder immune cells
Schmit et al. Cathespin K expression and actvitity in canine OSA
CatK is lysosomal protease with collagenolytic activity, and secretion by osteocalts is responsible for degrading bone matrix
Canine OS cells contained ___ CatK in cytoplasmic vesicles
In OS cells, ____ induced secretion CatK, which degraded ___
CatK concentration in OS compared to healthy dogs?
In subset of dogs, CatK decreased after palliatve RT and antiresorptive treatment
preformed
TGFB1, collagen type I
higher
Shoeneman et al. 2011. Expression and function of survivin in canine OSA
Elevated survivin correlated with?
Survivin attenutation lead to?
Increased histologic grade and MI and decreased DFI
Inhibited cell cycle progression, increased apoptosis, mitotic arrest, and chemosensitivity, and cooperated with chemotherapy to improve tumor control
Marley et al. The effects of taurolidine alone and in combination with doxorubicin or carboplatin in canine OSA in vitro
Taurolidine is derivative of taurine and has anti-tumor and anti-angiogenic effects against variety of cancers
Effect on OSA cells and in combination with dox or carbo?
Taurolidine cytotoxicity was ____ in 1 cell line
Effects may depend on functional status of p53
Cytotoxic and increased toxicity of dox and carbo in vitro
Caspace dependent with mutant p53 protein
Kruse et al. Evaluation of clinical and histopathologic prognostic factors for survival of canine OSA of extracranial flat and irregular bones
What does not appear to be correlated with survival?
What was associated with decreased survival time?
Histologic grade
Increased ALP and tumor location in scapula was associated with decreased survival times
Wells et al. Arginase treatment prevents the recovery of canine LSA and OSA cells resistant to the toxic effects of prolonged arginine deprivation
Effects of arginine deprication in canine LSA and OSA cell lines
Cell lines unable to recover 6 days of arginine deprivation but full recovery on return to arginine rich culture
Cell death after arginase
Adult progenitor cells from BM of healthy dog able to recover after 9 days of culture in arginase
Over 3 days in culture, arginase was more effective than asparaginase in inducing cell death in lymphoid cell lines
Kozicki et al. Adjuvant therapy with carboplatin and pamidronate for canine appendicular OSA
17 dogs treated with amputation
Median DFI compared to historical controls?
MST?
Addition of pamidronate to carboplatin is safe and does not impair efficacy of carboplatin treatment
185 days (6 mo) vs. 172 days
311 (10 mo) vs. 294 days (9.8 mo)
Sacornrattana et al. Abdominal ultrasonographic findings at diagnosis of OSA in dogs and association with treatment outcomes
How many had abnormalties?
Organ with most changes?
Metastases identified in how may dogs?
Dogs with abnormalties were less likley to receive definitive therapy, exhibited shorter survival (latter not significant)
Identifications of lesions in liver or kidney were statistically associated with shorter survival
57%
Spleen
2.5%
Cannon et al. Canine OSA cells exhibit resistance to aurora kinase inhibitors
Effects of aurora kinase inhibitors AZD1152 and VX680 on OSA cells
Cytotoxicity was observed but inhibitor concentrastion were higher in human leukemia and canine LSA cells
AZD1152 reduced aurora kinase phophorylation - resisatnce not due to target recognition
Efflux mediated by ABCB1 and ABCG2 was the not the mechanism
Marley et al. 2013. PK study and evaluation of safety of taurolidine for dogs with OSA
Taurolidine infusion safe in 6 dogs
Taurolidine with PVP (polyvinylpyrrolidine) had allergic reaction but recovered when infusion was stopped
3 dogs with OSA received dox and taurolidine - DCM, PLN, renal insufficiency, vasculopathy
1 dog develop ototoxicity with carbo and taurolidine
GI and BM toxicities were not increased over dox and carbo alone
Combo with doxo not recommend but with carbo appears safe
Stoewen et al. Factors influencing veterinarian referral to oncology specialists for treatment of dogs with LSA and OSA in Ontario, Canada
- 3% recommended referral for LSA and OSA
- 3% practised <2hr, 94% were confident in oncology service
22-33% were experienced with use of chemotherapeutics. 60% were experienced with amputation
Referral was associated with practioners perception of health status, interaction between clients bond with the dogs and clients financial status, practioner experience with treating cancer, how worthwhile practioner consider treatment to be, and confidence in referral center
Caro et al. 2013. Markers of ironmetabolism in retired racing urinse Greyhounds with and without OSA
Milovancev et al. Comparative analysis of the surface exposed proteome of 2 canine OSA cell lines and normal canine osteoblasts
Wallace et al. 2013. Diagnostic utility of AUS for routine staging at diagnosis of skeletal OSA in dogs
23 abnormalites were evaluated and 19 were bening
5% had another primary neoplasia
2/9 had AUS due to palpable abdominal mass was diagnosed with another neoplasia compared with only 1 of the 49 that had US for routine staging
Do US in animals with palpable abdominal mass - influence treatment decisions
Scharf et al. Effect of bevacizumab and growth of canine OSA cells xenografted in athymic mice
Mice in high-dose bevacizumab had significantly delayed tumor grwoth compared to low-dose group
VEGF inhibitors may be useful in treatment of OSA in dogs
Schwartz et al. 2013. Orthotopic model of canine OSA in athymic rats for evaluation of SRT
Tumors developed in 10/12 tibial sites and 14 femoral sutes
Administration of SRT was feasible
Mean tumor necrosis 95% and minimal AE on skin
Oblak et al. 2015. Comparison of concurrent imaging modalities for staging of dogs with appendicular primary bone tumors
No definitive lesions were identified on radiographs or CT
Lesions were identified on thoracic CT that were not visible radiographically
Ground glass pulmonary lesions progressed in 3/4 cases - suspicious for metastasis
Whole body CT was not suitable alternative to bone scintigraphy however useful as adjunctive diagnostic modality
Covey et al. Stereotactic radiosurgery and fracture fixation in 6 dogs with appendicular OSA
6 dogs
2 had pathologic fracture at admision and 4 after SRS
First 3 fractures repaired using open approach and 3 using minimally invasive percutaneous osteosynthesis
Infection occured in 5 dogs (83%) and implant failure in 3 (50%)
Survival times ranged from 364-897 days (1-2.5 yr)
Pang et al. Global gene expression analysis of canine osteosarcoma stem cells reveals a novel role for COX2 in tumor initiation
COX-2 was expressed 141-fold more in CSC than daughter adherent cells
COX-2 inhibition had no effect on CSC growth, or resistance to chemotherapy however it inhibited COX-2 in daughter cells prevented sphere formation - potential significance of COX-2 in tumor initiation
Selmic et al. Outcome and prognostic factors for OSA of the maxilla, mandible, calvarium in dogs: 183 cases
Mean age, Body weight, Breed?
Local progression occured in? How many developed metastatic disease
MST
Dogs that underwent surgery MST?
Tumor free margins were associated with decreased hazard of progression or recurrence
Dogs with OSA of calvarium had significantly greater hazard of local recurrence or progression
9.3 years, 32kg, purebred 44%
51%, 40%
239 days (8 mo)
329 days (11 mo)
Maniscalco et al. 2014. Increased expression of insulin life growth factor-1 receptor is correlated with worse survival in canine appendicular OSA
IGF-1R was expressed in ___ of OSA samples and dogs with higher levels had ____
IGF-1R is activated by ___, in paracrine or endocrine manner - activation of AKT/MAPK signaling, PPP caused p-IGF-1R dephosphorylation with partial blocking of p-MAPK and p-AKT, and apoptosis
IGF-1R expression is correlated with poor prognosis and potential therapeutic target
71%, decreased survival
IGF-1
Selmic et al. 2014. OSA following tibial plateau leveling osteotomy in dogs: 29 cases
Mean age, weight?
Associated clinical signs?
Mean interval between TPLO and OSA diagnosis?
Cast stainless steel in 18 dods and plates of wrough stainless steel in 4
MST for 10 dogs that underwent treatment with amputation and chemo?
Similar to outcomes reported for dogs with appendicular OSA
9 yr, 45 kg
Swelling 17/21 and lameness 28/29
5.3 years
313 days
Rodrigues et al. Tumorigenic canine OSA cell lines associated with frizzled-6 upregulation and enhanced population cell frquency
____ was upregulated in tumorigenic cell lines (7.78 fold) compared to non-tumorigenic cell lines
Frizzled-6, coreceptor for Wnt
Tumorigenic cell lines had increase in side population of cells compared with non-tumorigenic (6% vs. 1.6%)
There was no difference in tumorigenicity, frizzled-6, or percentage of side population cells noted b/t OSA cells lines generated from patient with differing ALP concentrations
Frizzled-6
Shor et al. 2015. Expression of nociceptive ligands in canine OSA
10 dogs with appendicular OSA
Canine OS cells express and secrete?
OS samples express?
Circulating noceiceptive ligand concentration were detectable but failed to correlate with pain status
NGF, endothelin-1, and PGE2
Noceiceptive ligands
Nagamine et al. 2015. Diversity of histologic patterns and expression of cytoskeletal proteins in canine skeletal OSA
64% of OS contained multiple subtypes
Myxoid, round cell, epitheliod subtypes were observed
Epitheloid were observed in OS of the head (OSH)
CK was expressed by epitheliod and sarcomatoid tumor cells, and all CK+ tumor cells expressed vimentin
Vimentin and SMA were expressed in all subtypes
GFAP was expressed in many subtypes, and some showed nerofilament expression
Mauchle et al. 2014. Identification of anti-proliferative kinase inhibitors as potential therapeutic agents to treat canine OSA
22 different kinase inhibitors
4 compounds (RO 31-8220, 5-iodorubericidin, BAY 11-7082, and erstatin) whoed significant cell growth inhibitor effect
RO 31-8220 and 5-iodorubericidin showed the highest ability to potentiate effects of doxorubicin
Targeting PKC, CK1, PKA, ErbB2, mTOR and NF-KB pathways
Sabattini et al. 2017. Comparative assessment of the accuracy of cytological and histologic biopsies in the diagnosis of canine bone lesions
Accuracy for cytology? Sensitivty and specificity
Accuracy for histology? Sensitivty and specificity
Tumor was correctly identified cytologically and histologically in?
83% - 83% and 80%
82.1% - 72% and 100%
50% and 55%
Tuohy et al. 2016. Association of canine OSA and monocyte phenotype and chemotactic function
Percentage of expression of CCR2 and CXCR2 was higher in control dogs compared to OSA
PGE2 and TNFa are increased in OSA monocyte culture compared to controls
Monocytes from OSA had decreased chemotactic function compared to control dogs
Decreased chemokine receptor expression and chemotaxis - evade immune response
Story et al. 2017. Evaluation of weight change during carboplatin therapy in dogs with appendicular OSA
A slight ___ in weight occured over the course of chemotherapy - but not statistically significant
Affect on survival?
Increase
Weight change did not have affect on survival
Byrum et al. 2016. Downregulation of CXCR4 expression and functionality after zoledronate exposure in canine OSA
20 dogs with OSA
All canine OSA cells express CXCR4 and zoledronate ___ expression and functionality by 27% through proteosome degradation and reduced prenylation of heterotrimeric G-proteins in 33% of tumor cell lines
In OS dogs, zoledronate reduces CXCR4 expression by 40% and decreases circulating CXCR4 in 18 of 20 dogs
Reduced
Burton et al. 2015. Implant associated neoplasia in dogs: 16 cases
Median time from implant to diagnosis of neoplasia?
Which limbs were most frquently affected?
Most common location on bone?
Most common tumor?
5.5 years
Hindlimbs - tibia 8/16, femur 5/16, 1 pelvis and femur
Diaphysis 15/16
OSA 13/16, HS, FSA, spindle cell sarcoma
Rubin et al. Factors associated with pathological fractures in dogs with appendicular primary bone neoplasia: 84 cases
Median time from diagnosis to euthanasia?
Pathological fractures identified in how many dogs?
Most commonly affcted bones?
Tumors arising from long bones other than ___ had odds of eventual fracture 5.05 as great as odds for tumors of the radius
Lytic tumors had odds eventual fracture 3.22 as great as odds for tumors that appeared blastic or mixed lytic-blastic
Radial tumors less likely to fracture and lytic tumors more likely to fracture
111 days
38%
Femur>tibia>humerus>radius>ulna
Radius
Meyer et al. 2015. Expression of platelet-derived growth factor BB, erythropoeitin and erythropoeiting receptor in canine and feline OSA
EPO-R is expressed on neoplastic cells - may play a role in tumor progression
EPO expression was positive in all tumors and EPO mRNA was detected in 38% of canine and 40% feline samples
EPO-R was expressed in all samples
EPO-R mRNA was higher in feline tumors, tumor cell lines, and kidney compared to canine tissues
All cell lines responded to EPO treatment
Autocrine and paracrine tumor progression cannot be excluded
Willcox et al. Serum 25-hydroxyvitamin D concentrations in dogs with OSA do not differ from those age- and weight- matched control dogs
Levels were similar in dogs with OSA and matched controls
25-hydroxyvitamin-D insufficiency may not play a role in pathogenesis
Zhang et al. 2015. The effect of Zhangfei/CREBZF on cell growth, differentiation, apoptosis, migration, and the unfolded protein response in several canine OSA cell lines
Like D-17 the 3 cell lines expressed p53 proteins capable of activating with p53 response elements
Zhangfei suppressed the growth of UPR-related transcripts
Induced activation of osteocalcin expression a marker of osteoblast differentiation and trigged programmed cell death
Induction of Zhangfei, where p53 is functional, may be an effective modality for treatment of OS
Herrera et al. 2015. Peroxisome proliferator activated receptor g protein expression is assymmetrically distributed in primary lung tumor and metastatic to lung OSA samples and does not correlate with gene methylation
PPAR-g is a TF plays a role in proliferation and differentiation
PPAR-g is expressed in __% of canine NSCLC and __ % of metastatic OSA
There was significant loss of 5’PPAR-g methylation from normal lung to primary lung cancer to metastatic OSA
Altered PPAR-g promoter methylation at the interrogated locus does not appear to be associated with changes in protein expression
33%, 66%
Fowles et al. Intra- and interspecies gene expression models for predicting drug response in canine OSA
Mantovani et al. Effects of epidermal growth factor receptor kinase inhibiton on radiation response in canine OSA cells
Effects of small molecule EGFR inhibitor erlotinib on canine OSA radiation response
Erlotinib reduced clonogenic survival in 2 canine OSA cell lines and enhacned the impact of RT in 1/3 cell lines
Erlotinib enhanced radiation effects and demonstrated single effects
Did not alter EGFR nor inhibit PKB/Akt - increased phoshphorylation of Akt
VEGF levels increased after erlotinib as single agent and in combination with RT in 2/3 and decreased in 1/3 cell lines
Levine et al. 2016. Effects and synergy of feed ingredients on canine neoplastic cell proliferation
Examine 5 feed ingredients for antiproliferative effects and interaction with toceranib and doxorubicin
Which agents were most effective?
___ extract was most potent and exhibited synergy with __ extract
Combination had additive or synergistic effects with chemo agents
Tea, turmeric, and rosemary extract
Turmeric, rosemary
Murphy et al. 2017. Evaluation of P16 expression in canine appendicular OSA
Lack of canine OSA P16 expression and shorter disease free interval
Ong et al. Anti-tumor efficacy of etoposide alone and in combination with piroxicam against canine OSA in xenograft model
Etoposide single agent delayed tumor progression with marked reduction of Ki67 in tumor tissue
Combination treatment (with piroxicam) did not enhance anti-tumor efficacy of etoposide
Both (etoposide alone and combination) downregulated survivin expression but was not followed by increased apoptosis
Leeper et al. Preliminary evaluation of serum total cholesterol concentrations in dogs with OSA
Total cholesterol was ___ in OSA bearing dogs, ___% fracture controls, and __% similar age/weight controls
Elevated cholesterol was associated with?
elevated 45%, 10%, 6.5%
reduced hazard ratio for overall mortality in dogs with OSA
Kubicek et al. Association between CT characteristics and fractures following stereotactic radiosurgery in dogs with appendicular OSA
MST?
Fracture free rates at 3, 6 and 9 month?
Bone affected was significantly associated with time to fracture
Median time to fracture in dogs with subchondral bone involvement? without subchondral bone involvement?
Acute and late skin effects 58% and 16%
9.7 months
73%, 44%, 38%
4.2 months, 16.3 months
Pagano et al. Safety and toxicity of an accelerated coarsely fractionated radiation protocol for treatment of appendicular OSA in 14 dogs
10Gy delievered in 2 consecutive days - total of 20Gy
13/14 received adjuvant therapy with pamidronate and NSAID
9 dogs received chemotherapy
Median of __ days, % had improved pain control
Median DOR?
How many developed pathologic fracture?
Timing of fracture?
14 days, 93%
80 days
35.7%
24-250 days
Arthur et al. Risk of OSA in dogs after open fracture fixation
19,041 fractures, 15 dogs developed OSA affecting the same bone
Seguin et al. Long term outcome of dogs treated with ulnar rollover transposition for limb-sparing of distal radial OSA: 27 limbs in 26 dogs
Complication occured in how many limbs?
Infection?
Biomechanical complication?
Local recurrence?
Limb function graded by veterinarians or owner was poor in 2 limbs, fair in 4, good in 15, excellent in 3, unknown iun 4
Median DFI and ST
High complication rate but limb function was acceptable
20 (76%)
12 (48%)
15 (58%
2 (8%)
245 (8 mo) and 277 days (9 mo)
BMI1 is expressed in canine OSA and contributes to cell growth and chemotherapy resistance
BMI1 is stem cell factor
BMI1 expression in primary and metastatic OSA?
Canine OSA cell lines expressed high levels of BMI1 compared with osteoblasts and knockdown demonstrated resistance to carboplatin and doxorubicin chemotherapy
Inhibition of BMI1 may improve response to chemotherapy
Strong nuclear expression
Wehrle-Martinez et al. Osteocalcin and osteonectin expression in canine OSA
23 OSA, 4 CSA, 4 FSA, 2 HSA, 4 HS
OC expression in OSA and CSA?
OC in FSA, HSA, HS?
sensitivity and specificity for OC in canine OSA?
ON?
83% OSA and 100% CSA
Negative
83% and 71%
100% OSA and 100% non-OSA - 100% sensitivty but no specificty
Massimi et al. 17-AAG and apoptosis, autophagy, and mitophagy in canine OSA cell lines
Roy et al. Comparative proteomic investigation of metastatic and non-metastatic osteosarcoma cells of human and canine origin
Rebhun et al. Evaluation of optimal water flouridation on the incidence and skeletal distribution of naturally arising OSA in pet dogs
No difference in age, skeletal distrubtion of OSA cases relative to flouride status
Dogs with OSA were not more likely to live in area with optimized flouride in water than dogs with LSA or HSA
Does not contribute to OSA
Rizzo et al. The effects of sulforaphance on canine OSA proliferation and invasion
SFN found in raw cruciferous vegetables may have utility in chemoprevention - as antineoplastic and free radical scavenger
SFN could not induce cell death at physiologic concentrations but diminished cell invasion and downregulation of focal adhesion kianse (FAK) signaling
When used with doxorubicin there was a protective effect to dox-induced cytotoxicity
York et al. Enrofloxacin enhances the effects of chemotherapy in canine OSA cells with mutant and wild-type p53
Moresco and Abrams contained mutations in p53 while no mutation in D17 cell line or normal canine osteoblast cell line
The addition of enrofloxacin to either dox or carbo resulted in further reduction in cell viability - regardless of p53 mutational status
Guiffrida et al. Use of routine histopathology and factor VIII-related antigen/vWF IHS to differentiate primary HSA of bone from telengiectatic OSA in 54 dogs
20% of tumors were reclassified based on FVIII-RAg/vWF immunoreactivity from original diagnosis of tOSA
Laver et al. Prospective evaluation of toceranib phosphate in metastatic canine OSA
AEs?
SD?
Median PFS
Plasma VEGF concentration were significantly elevated at 4 weeks of TOC treatment, but no changes in Treg
Common but low grade
18% with remainder having PD
89 days
Wouda et al. 2017. Safety evaluation of combination carboplatin and toceranib phosphate in tumor bearing dogs: A phase I dose finding study
MTD of carboplatin?
Palladia?
Dose limiting toxicity?
Clinical benefit?
200 mg/m2 IV q 21 days
2.75 mg/kg PO EOD
neutropenia
Observed in most cases
Bulla et al. 2017. Platelets inhibit migration of canine OSA cells
Addition of blood leukocytes to platelet samples did not alter the inhibitory effect on migration
Platelet treatment downregulated?
Interaction between canine platelets or molecules released during platelet activation inhibits their migration - canine pletelets may antagonize metastasis of OSA
SNAI2 and TWIST1 genes - genes involved in EMT
Frimberger et al. Canine OSA treated by post-amputaion sequential accelerated doxorubicin and carboplatin chemotherapy: 38 cases
MST? 1- and 2- year survival
How many dogs hospitalized?
What impacted survival time on multivariate analysis?
Outcomes were similar to other chemo protocols
317 days (10.5 mo), 43% and 14%
5.2%
Serum ALP and whether chemo was completed
Goupil et al. 2012. Prevelence of 5-lipooxygenase expression in canine OSA and the effects of dual 5-LOX/COX inhibitor on OSA cells in vitro and in vivo
5-LOX inhibitor, tepoxalin, reduced cell proliferation
5-LOX leads to production of lipid mediations and when added to media did not recover cell proliferation
Lipid mediation production is not the means by which tepoxalin alters proliferation
Montinaro et al. 2013. Clinical outcome of 42 dogs with scapular tumors treated by scapulectomy: VSSO retrospective study
Subtotal scapulectomy done in 18 dogs (43%)
Limb use was good to excellent overall
Only adjunctive chemotherapy had positive effect on DFI
London et al. 2015. Impact of toceranib/piroxicam/cyclophosphamide maintenace therapy on outcome of dogs with appendicular OSA follwoing amputation and carbopaltin chemotherapy
Median DFI for control and toceranib treated dogs was?
Median OS for control and toceranib?
1-year survival rate for control and toceranib?
Addition of toceranib to metronomic piroxicam/cyclophpsphamide therapy following amp + carbo did not improve DFI
215 (7 mo) and 233 days
242 (8 mo) and 318 days (10.6 mo)
35% and 38%
Mason et al. 2016. Immunotherapy with Her2-targeting listeria induces HER2-specific immunity and demonstrates potential therapeutic effects in a phase I trial in canine OSA
ADXS31-164 - attenutaed, Listeria monocytogenes expressing human HER2/neu fusion protein to induce immunity
Toxicities?
ADXS31-164 induced IFN-g response in 15/18 dogs within 6 months
ADXS31-164 reduced metastatic disease and increased duration of survival time
Low grade
Curtis et al. 2016. Combination therapy with zoledronic acid and PTH improves bone architecture and strenght following a clincially relevant dose of SRT for the local tretament of canine OSA in athymic rats
Significant increase in bone volume and increased polar moments of inertia 8 weeks after radiation in the combine ZA/PTH treatment group as compared to RT alone
Fenger et al. 2016. MIR-9 is overexpressed in spontaneous canine OSA and promotes a metastatic phenmotype inlcusing invasion and migration in osteoblats and OSA cell lines
Mir-9 is overexpressed in OS tumors and cell lines
Transduced normal osteoblasts and cell lines with mir9 - invasion and migration but did not affect proliferation or apoptosis
Overexpression of mir9 identified alterations in numerous genes - upregulation of GSN, an actin filament severing protein involved in cytoskeletal remodeling
Downregulation of mir-9 in OS cell lines reduced GSN with decrease in invasion and migration
Talbot et al. 2017. Retrospective evaluation of whole body CT for tumor staging in dogs with primary appendicular OSA
39 dogs
Was bone metastasis detected?
Pulmonary metastasis?
How many dogs had additional masses detected?
No
5%
5%
Ong et al. 2016. Effects of etoposide alone and in combination with piroxicam on canine OSA cell lines
Effects of etoposide alone on cell line?
Effects of etoposide in combination with piroxicam?
What caused tha anti-proliferative effect?
Supressed cell growth and viability
Concentration dependent cytotoxicity
Cdc2-CyclinB1 - increase in G2/M fraction
Boston et al. 2017. Outcome and complications in dogs with appendicular primary bone tumors treated with stereotactic radiotherapy and concurrent surgical stabilization.
18 dogs
7 dogs had pathological fracture and 11 were at high risk
single dose of SRT (5) or 3 doses (13)
Bone plate (15), interlocking nail (3)
Complications occured in how many dogs?
Most common complication?
Post-operative fracture was recored in how many?
9 dogs were amputated at median of?
MST?
Treatement with SRT and stabilization associated with high complication rate - consider alternative methods of limb salvage
16/17, 15/17 being major complicarions
Infection - 15/17
3 cases
152 days
344 days (11.5 mo)
Turner et al. 2017. Prognosis of dogs with stage III OSA following treatment with amputation and chemotherapy with and without metastasectomy
MST for dogs that received no treatment for metastasis?
MST for dogs that received metastatectomy and did not?
Basis used for metastasectomy?
2 months
232 days (7.7 mo) vs. 49 days
<3 pulmonary nodules on CXR and DFI>275 days
Al-Khan et al. 2017. Immunohistochemical validation of spontaneously arising canine OSA as a Model for Human OSA
Markers expressed in canine OSA?
Markers variably expressed in canine OSA?
Similar to human OSA, canine OSA may represent model to study the disease
Vimentin, ALP, Runx2 (runt-related TF4), BMP4 (bone morphogenetic protein 4)
Desmin, S100, NSE (neuron specific enolase)
Swift et al. 2018. Outcome of 9 dogs treated with SRT for primary or metastatic vertebral OSA
9 dogs treated with SRT - 1 to 5 fractions with total dose 13.5 - 36Gy
6 dogs primary lesions and 3 had metastatic lesions
Neurologic signs improved in 4 patients, remained the same in 4 and worsened in 1
5/6 dogs that presented with assessable spinal pain reported improvement in pain
MST?
Median duration of pain control?
139 days (4.6 mon)
77 days
Leonardo et al. 2017. miR-1 abd miR-133b expression in canine OSA
Important role in regulation osteogenesis and tumor cell proliferation
Expression in tumors compared to normal bones?
Expressionof MET and MCL1
Potential role of miR-1 and miR-133b as biomarkers
Lower expression in tumors compared to normal bone with higher expression of target genes MET and MCL1
Moderate to strong immunostaining in more than 50% tumor samples
Schott et al. 2018. Histologic grade does not predict outcome in dogs with appendicular OSA receiving the standard of care
Compare 2 published histologic grading systems with appendicular OSA treated with SOC for curative intent
Histologic grade and outcome?
What was correlated with outcome?
Histologic grade did not correlate with outcome
Increased number of mitotic figures per 3/400x
Selmic et al. 2018. Association of TPLO with proximal tibial OSA in dogs
TPLO had been performed >1year ago
Dog with TPLO were ___ to develop OSA than dogs without
Each 1kg increased in body weight was associated with 11% increase in odds of proximal tibibal OSA
OSA should be differential for new or worsening hindlimb lameness in dogs that underwent TPLO >1 year previously
40x
Matsuyama et al. 2018. Evaluation of metronomic cyclophosphamide chemotherapy as maintenance for dogs with appendicular OSA following limb amputation and carboplatin chemotherapy
Dogs grouped whether they received MC or did not (15 mg/m2 PO q24h and meloxicam 0.1 mg/kg PO Q24hr)
19 dogs received carboplatin and MC chemotherapy and 20 received carboplatin chemotherapy
Treatment discontinued in how many dogs? Cause?
Median follow up period?
Difference in MST and PFS between the 2 groups?
58% due to cystitis
450 days
None
Sprecher et al. 2018. Retrospective analysis of corrosion and ion release from retrieved cast stainless steel tibial plateu leveling osteotomy plates in dogs with and without pre-implant OSA
Greater metal ion release was observed in cast plates than foreged plates
Accumulation of metal ions may trigger neoplastic transformation in neighboring cells
Withers et al. 2018. Association of macrophages and lymphocyte infiltration with outcome in canine OSA
Median number of CD3+ and FOXP3+ infiltrates were 45.8 cells/mm2 and 8.5 mm2
The median area of CD204+ macrophages was 4.7% and dogs with tumors contained greater than 4.7% CD204+ experienced significantly longer DFI
Macrophages may play a role in inhibiting cOSA progression
Rici et al. 2018. Combination therapy of canine OSA with bone marrow stem cells, bone morphogenetic proteina and carboplatin in an in vivo model
Combination of mesenchymcal stem cells and rhBMP-2 reduced tumor mass and infiltrastion of neoplastic cells in tissues more efficiently than carboplatin alone
Patatsos et al. 2018. Pre-clionical evaluation of protesosome inhibitors for canine and human OSA
Gold et al. 2018. Zygomatic arch parosteal OSA in dogs and cats
Parosteal OSA is slow growing tumors arising from surface of long bones
5 dogs and 1 cat
Clinical presentation varied from chronic sneezing to facial swelling
Histologically - well differentiated fibro-osseous and chondroid components
Cellular atypia and mitotic figures were uncommon
Hans et al. 2018. Effect of surgical site infection on survival after limb amputation in the curative-intent treatment of canine appendicular OSA
15 dogs with OSA and SSI and 134 dogs with OSA and no SSI
Median DFI and MST
Difference in DFI dogs with SSI and without SSI?
MST with SSI and without SSI?
What was associated with decreased DFI and survival?
SSI did not influence survival
236 days (8 mo) and 283 days (9.4 mo)
Not different - 292 days with and 224 days without
Not different - 292 days and 280 days
Not completeing chemotheray
Guiffrida et al. 2018. Primary appendicular HSA and tOSA in 70 dogs
Dogs with HSA were more likely than dogs with tOSA to be ___ and have ___ limb tumors
__% of HSA occured in hind limbs, particularly ___
Dogs with tOSA weighed a median of 9.9kg more than dogs with HSA
OS for HSA and tOSA?
Younger age and more aggressive treatment were associated with longer survival in dogs with HSA but not tOSA
1 year survival rates did not differ between HSA (28%) abd tOSA (7%)
Males and hind limb tumors
78%, tibia
299 days (10 mon) and 213 days (7 mon)
