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
