QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
Osteosarcoma Publications
1. Li Y, Flores R, Yu A, Okcu MF, Murray J, Chintagumpala M, Hicks J, Lau CC, Man TK.
Elevated Expression of CXC Chemokines in Pediatric Osteosarcoma Patients. Cancer
117(1): 207–17, 2011 Jan 1.
http://dx.doi.org/10.1002/cncr.25563
Abstract
BACKGROUND:
Osteosarcoma is the most common malignant bone tumor in children. Despite the advent
of chemotherapy, the survival of osteosarcoma patients has not been significantly
improved recently. Chemokines are a group of signaling molecules that have been
implicated in tumorigenesis and metastasis.
METHODS:
The authors used an antibody microarray to identify chemokines that were elevated in the
plasma samples of osteosarcoma patients. The results were validated using enzymelinked immunosorbent assays on an independent set of samples. The tumor expressions
of 3 chemokines were examined in 2 sets of osteosarcoma tissue arrays. The authors
also evaluated the proliferative effect of the chemokines in 4 osteosarcoma cell lines.
RESULTS:
The authors found that the plasma levels of CXCL4, CXCL6, and CXCL12 in the
osteosarcoma patients were significantly higher than those in the controls, and the results
were validated by an independent osteosarcoma cohort (P < .05). However, CXCL4
(100%) and CXCL6 (91%) were frequently expressed in osteosarcoma, whereas CXCL12
was only expressed in 4%. Survival analysis further showed that higher circulating levels
of CXCL4 and CXCL6, but not CXCL12, were associated with a poorer outcome of
osteosarcoma patients. Addition of exogenous chemokines significantly promoted the
growth of different osteosarcoma cells (P < .05).
CONCLUSIONS:
The results demonstrate that CXCL4 and CXCL6 are frequently expressed in
osteosarcoma, and that the plasma levels of these 2 chemokines are associated with
patient outcomes. Further study of these circulating chemokines may provide a promising
approach for prognostication of osteosarcoma. Targeting these chemokines or their
receptors may also lead to a novel therapeutic invention. Cancer 2011. © 2010 American
Cancer Society.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
2. Janeway KA, Barkauskas DA, Krailo MD, Meyers PA, Schwartz CL, Ebb DH, Seibel NL, Grier
HE, Gorlick R, Marina N. Outcome for adolescent and young adult patients with
osteosarcoma: A Report from the Children’s Oncology Group. Cancer 118(18): 4597–4605.
http://dx.doi.org/10.1002/cncr.27414
Abstract
BACKGROUND:
There are conflicting data regarding age as a prognostic factor in osteosarcoma. The
authors conducted a study evaluating the impact of age on prognosis in children and
young adults with osteosarcoma enrolled on North American cooperative group trials.
METHODS:
Patients with high-grade osteosarcoma of any site enrolled on North American
cooperative group trials CCG-7943, POG-9754, INT-0133, and AOST0121 were included
in this study. Primary tumor site, age, sex, ethnicity, histologic response, and presence of
metastatic disease at diagnosis were evaluated for their impact on overall survival (OS)
and event-free survival (EFS).
RESULTS:
A total of 1054 patients were eligible and had complete data available for the study. Age
was not significantly associated with any other presenting covariate analyzed except sex.
Age 18 or older was associated with a statistically significant poorer EFS (P = .019) and
OS (P = .043). The 10-year EFS and OS in patients <10, 10 to 17, and ≥18 years old
were 55%, 55%, 37% and 68%, 60%, 41%, respectively. The poorer EFS in patients ≥18
years old was because of an increased rate of relapse. Presence of metastatic disease at
diagnosis, poor histologic response, and pelvic tumor site were also associated with a
poorer prognosis. In multivariate analysis, age continued to be associated with poorer
EFS (P = .019) and OS (P = .049).
CONCLUSIONS:
In osteosarcoma, age 18 to 30 years is associated with a statistically significant poorer
outcome because of an increased rate of relapse. Poorer outcome in adolescent and
young adult patients is not explained by tumor location, histologic response, or metastatic
disease at presentation. Cancer 2012. © 2012 American Cancer Society.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
3. Isakoff MS, Barkauskas DA, Ebb DH, Morris CD, Letson GD. Poor Survival for Osteosarcoma
of the Pelvis: A report from the Children’s Oncology Group. Clinical Orthopaedics and
Related Research 470(7): 2007–2013.
http://dx.doi.org/10.1007/s11999-012-2284-9
Abstract
Background
The pelvis is an infrequent site of osteosarcoma and treatment requires surgery plus
systemic chemotherapy. Poor survival has been reported, but has not been confirmed
previously by the Children’s Oncology Group (COG). In addition, survival of patients with
pelvic osteosarcomas has not been compared directly with that of patients with nonpelvic
disease treated on the same clinical trials.
Questions/purposes
First, we assessed the event-free (EFS) and overall survival (OS) of patients with pelvic
osteosarcoma treated on COG clinical trials. We then asked whether patient survival
compared with that of patients treated on the same clinical trials with nonpelvic disease.
Finally, we asked whether patients with metastatic disease at initial diagnosis had worse
survival.
Methods
We retrospectively reviewed data from 1054 patients with osteosarcoma treated in four
studies between 1993 and 2005. Twenty-six of the 1054 patients (2.5%) had a primary
tumor of the pelvis. At diagnosis, nine patients had metastatic disease. The minimum
followup was 2 months (mean, 34 months; range, 2–102 months).
Results
Two of the nine patients with metastatic disease at diagnosis and five of the 17 with
localized disease were alive at last contact. Estimates of the 5-year EFS for localized
versus metastatic disease of the pelvis were 22% versus 23%. OS for patients with
localized versus metastatic disease was 47% versus 22%. Patients with osteosarcoma in
all other locations had a 5-year EFS of 57% and OS of 69%.
Conclusions
Our analysis confirms poor survival for patients with pelvic osteosarcoma. Survival with
metastatic disease in the absence of a pelvic primary tumor is similar to that for localized
or metastatic pelvic osteosarcoma. Improved surgical or medical therapy is needed, and
patients with pelvic osteosarcoma may warrant alternate or experimental therapy.
Level of Evidence
Level II, prognostic study. See Guidelines for Authors for a complete description of levels
of evidence.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
4. Jacobs KB, et al. Detectable clonal mosaicism and its relationship to aging and cancer.
Nature Genetics 44(6): 651–658.
http://dx.doi.org/10.1002/cncr.26112
Abstract
In an analysis of 31,717 cancer cases and 26,136 cancer-free controls from 13 genomewide association studies, we observed large chromosomal abnormalities in a subset of
clones in DNA obtained from blood or buccal samples. We observed mosaic
abnormalities, either aneuploidy or copy-neutral loss of heterozygosity, of >2 Mb in size in
autosomes of 517 individuals (0.89%), with abnormal cell proportions of between 7% and
95%. In cancer-free individuals, frequency increased with age, from 0.23% under 50
years to 1.91% between 75 and 79 years (P = 4.8 × 10(-8)). Mosaic abnormalities were
more frequent in individuals with solid tumors (0.97% versus 0.74% in cancer-free
individuals; odds ratio (OR) = 1.25; P = 0.016), with stronger association with cases who
had DNA collected before diagnosis or treatment (OR = 1.45; P = 0.0005). Detectable
mosaicism was also more common in individuals for whom DNA was collected at least 1
year before diagnosis with leukemia compared to cancer-free individuals (OR = 35.4; P =
3.8 × 10(-11)). These findings underscore the time-dependent nature of somatic events in
the etiology of cancer and potentially other late-onset diseases.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
5. Ebb D, Meyers P, Grier H, Bernstein M, Gorlick R, Lipshultz SE, Krailo M, Devidas M,
Barkauskas DA, Siegal GP, Ferguson WS, Letson GD, Marcus K, Goorin A, Beardsley P,
Marina N. (2012) Phase II Trial of Trastuzumab in Combination With Cytotoxic
Chemotherapy for Treatment of Metastatic Osteosarcoma With Human Epidermal
Growth Factor Receptor 2 Overexpression: A Report From the Children's Oncology Group.
JCO 30(20): 2545–2551.
http://dx.doi.org/10.1200/JCO.2011.37.4546
Abstract
Purpose Despite efforts to intensify chemotherapy, survival for patients with metastatic
osteosarcoma remains poor. Overexpression of human epidermal growth factor receptor
2 (HER2) in osteosarcoma has been shown to predict poor therapeutic response and
decreased survival. This study tests the safety and feasibility of delivering biologically
targeted therapy by combining trastuzumab with standard chemotherapy in patients with
metastatic osteosarcoma and HER2 overexpression.
Patients and Methods Among 96 evaluable patients with newly diagnosed metastatic
osteosarcoma, 41 had tumors that were HER2-positive by immunohistochemistry. All
patients received chemotherapy with cisplatin, doxorubicin, methotrexate, ifosfamide, and
etoposide. Dexrazoxane was administered with doxorubicin to minimize the risk of
cardiotoxicity from treatment with trastuzumab and anthracycline. Only patients with
HER2 overexpression received concurrent therapy with trastuzumab given for 34
consecutive weeks.
Results The 30-month event-free and overall survival rates for patients with HER2
overexpression treated with chemotherapy and trastuzumab were 32% and 59%,
respectively. For patients without HER2 overexpression, treated with chemotherapy
alone, the 30-month event-free and overall survival rates were 32% and 50%,
respectively. There was no clinically significant short-term cardiotoxicity in patients treated
with trastuzumab and doxorubicin.
Conclusion Despite intensive chemotherapy plus trastuzumab for patients with HER2positive disease, the outcome for all patients was poor, with no significant difference
between the HER2-positive and HER2-negative groups. Although our findings suggest
that trastuzumab can be safely delivered in combination with anthracycline-based
chemotherapy and dexrazoxane, its therapeutic benefit remains uncertain. Definitive
assessment of trastuzumab's potential role in treating osteosarcoma would require a
randomized study of patients with HER2-positive disease.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
6. Savage SA, et al. Genome-wide Association Study Identifies Novel Loci Associated with
Osteosarcoma. Nature Genetics 45(7): 799–803.
http://dx.doi.org/10.1038/ng.2645
Abstract
Osteosarcoma is the most common primary bone malignancy of adolescents and young
adults. In order to better understand the genetic etiology of osteosarcoma, we performed
a multi-stage genome-wide association study (GWAS) consisting of 941 cases and 3,291
cancer-free adult controls of European ancestry. Two loci achieved genome-wide
significance: rs1906953 at 6p21.3, in the glutamate receptor metabotropic 4 [GRM4] gene
(P = 8.1 ×10-9), and rs7591996 and rs10208273 in a gene desert on 2p25.2 (P = 1.0
×10-8 and 2.9 ×10-7). These two susceptibility loci warrant further exploration to uncover
the biological mechanisms underlying susceptibility to osteosarcoma.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
7. Altaf S, Enders F, Jeavons E, Krailo M, Barkauskas DA, Meyers P, Arndt C.High BMI at
diagnosis is associated with inferior survival in patients with osteosarcoma: A report from
the Children’s Oncology Group. Pediatric Blood & Cancer 60(12): 2042–2046.
http://dx.doi.org/10.1002/pbc.24580
Abstract
BACKGROUND:
Body mass index (BMI), at diagnosis has been associated with lower survival and
increased toxicity in cancer patients. We analyzed the effect of BMI at diagnosis on
therapy related toxicities and outcome in pediatric osteosarcoma patients treated on
Children's Oncology Group (COG) trial INT0133.
PROCEDURES:
All patients enrolled on COG-INT0133 with height, weight and toxicity information were
eligible. BMI was expressed as age and gender specific percentiles using height and
weight at diagnosis. Patients were classified into high, normal and low BMI groups.
Logistic regression models were used to analyze toxicities; Kaplan-Meier curves were
created to assess event free (EFS) and overall survival (OAS).
RESULTS:
Seven hundred and ten patients met eligibility criteria. BMI distribution was: 447 normal
BMI, 74 low BMI, and 189 high BMI. Renal toxicity was higher in the high BMI group (OR
= 2.7, 95% CI 1.2-6.4, P = = 0.01) only during one of the courses of therapy. Compared to
the normal BMI group, patients with high BMI had significantly worse OAS at 5 years
compared to those with normal BMI, 69.7% versus 80.5% (HR = 1.6, 95% CI 1.1-2.2, P =
0.005) and a trend towards worse event-free survival at 3 years 66.2% versus 75.5% (HR
= 1.3 95% CI 0.9-1.8, P = 0.05). There was no difference in EFS or OAS in patients with
low BMI compared to patients with normal BMI.
CONCLUSIONS:
High BMI at diagnosis is associated with worse OAS in patients with osteosarcoma. No
clinically significant differences in toxicity were observed in the various BMI groups.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
8. Borinstein SC, Barkauskas DA, Bernstein M, Goorin A, Gorlick R, Krailo M, Schwartz
CL, Wexler LH, Toretsky JA. Analysis of serum insulin growth factor-1 concentrations in
localized osteosarcoma: A children’s oncology group study. Pediatric Blood & Cancer
61(4): 749–752.
http://dx.doi.org/10.1002/pbc.24778
Abstract
To investigate the role of insulin-like growth factor-1 (IGF-1), in localized osteosarcoma,
serum levels of IGF-1, IGFBP-2, and IGFBP-3 were measured in 224 similarly treated,
newly diagnosed patients. We demonstrated that younger patients had lower
concentrations of IGF-1 and IGFBP-3 compared to older (P < 0.001) along with lower
IGFBP-3:IGF-1 and IGFBP-2:IGF-1 ratios (P < 0.001). IGFBP-2 did not correlate with age
(P = 0.16), yet IGFBP-2:IGF-1 ratios were higher in the younger population (P < 0.001).
These findings show that older patients have higher concentrations of free IGF-1. None of
IGF-1, IGFBP-2, nor IGFBP-3 concentrations were associated with event-free nor overall
survival.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
9. Gorlick S, Barkauskas DA, Krailo M, Piperdi S, Sowers R, Gill J, Geller D, Randall RL,
Janeway K, Schwartz C, Grier H, Meyers PA, Gorlick R, Bernstein M, Marina N. (2014) HER2 expression is not prognostic in osteosarcoma; a Children's Oncology Group prospective
biology study. Pediatric Blood & Cancer 61(9): 1558–1564.
http://dx.doi.org/10.1002/pbc.25074
Abstract
Background
Since the initial reports of human epidermal growth factor receptor 2 (HER-2) expression
as being prognostic in osteosarcoma, numerous small studies varying in the interpretation
of the immunohistochemical (IHC) staining patterns have produced conflicting results.
The Children's Oncology Group therefore embarked on a prospective biology study in a
larger sample of patients to define in osteosarcoma the prognostic value of HER-2
expression using the methodology employed in the initial North American study
describing an association between HER-2 expression and outcome.
Procedure
The analytic patient population was comprised of 149 patients with newly diagnosed
osteosarcoma, 135 with localized disease and 14 with metastatic disease, all of whom
had follow up clinical data. Paraffin embedded material from the diagnostic biopsy was
stained with CB11 antibody and scored by two independent observers. Correlation of
HER-2 IHC score and demographic variables was analyzed using a Fisher's exact test
and correlation with survival using a Kaplan–Meier analysis.
Results
No association was found with HER-2 status and any of the demographic variables tested
including the presence or absence of metastatic disease at diagnosis. No association was
found between HER-2 status and either event free survival or overall survival in the
patients with localized disease.
Conclusion
HER-2 expression is not prognostic in osteosarcoma in the context of this large
prospective study. HER-2 expression cannot be used as a basis for stratification of
therapy. Identification of potential prognostic factors should occur in the context of large
multi-institutional biology studies. Pediatr Blood Cancer 2014;
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
10. Khanna C, et al. Towards a Drug Development Path that Targets Metastatic Progression in
Osteosarcoma. Clinical Cancer Research 20(16): 4200-4209.
http://dx.doi.org/10.1158/1078-0432.CCR-13-2574
Abstract
Despite successful primary tumor treatment, the development of pulmonary metastasis
continues to be the most common cause of mortality in patients with osteosarcoma. A
conventional drug development path requiring drugs to induce regression of established
lesions has not led to improvements for patients with osteosarcoma in more than 30
years. On the basis of our growing understanding of metastasis biology, it is now
reasonable and essential that we focus on developing therapeutics that target metastatic
progression. To advance this agenda, a meeting of key opinion leaders and experts in the
metastasis and osteosarcoma communities was convened in Bethesda, Maryland. The
goal of this meeting was to provide a “Perspective” that would establish a preclinical
translational path that could support the early evaluation of potential therapeutic agents
that uniquely target the metastatic phenotype. Although focused on osteosarcoma, the
need for this perspective is shared among many cancer types. The consensus achieved
from the meeting included the following: the biology of metastatic progression is
associated with metastasis-specific targets/processes that may not influence grossly
detectable lesions; targeting of metastasis-specific processes is feasible; rigorous
preclinical data are needed to support translation of metastasis-specific agents into
human trials where regression of measurable disease is not an expected outcome;
preclinical data should include an understanding of mechanism of action, validation of
pharmacodynamic markers of effective exposure and response, the use of several murine
models of effectiveness, and where feasible the inclusion of the dog with naturally
occurring osteosarcoma to define the activity of new drugs in the micrometastatic disease
setting. Clin Cancer Res;
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
11. Wang Z, et al. (2014) Imputation and subset based association analysis across different
cancer types identifies multiple independent risk loci in the TERT-CLPTM1L region on
chromosome 5p15.33. Human Molecular Genetics 23(24): 6616–6633.
http://dx.doi.org/10.1093/hmg/ddu363
Abstract t
Genome-wide association studies (GWAS) have mapped risk alleles for at least 10
distinct cancers to a small region of 63 000 bp on chromosome 5p15.33. This region
harbors the TERT and CLPTM1L genes; the former encodes the catalytic subunit of
telomerase reverse transcriptase and the latter may play a role in apoptosis. To
investigate further the genetic architecture of common susceptibility alleles in this region,
we conducted an agnostic subset-based meta-analysis (association analysis based on
subsets) across six distinct cancers in 34 248 cases and 45 036 controls. Based on
sequential conditional analysis, we identified as many as six independent risk loci marked
by common single-nucleotide polymorphisms: five in the TERT gene (Region 1:
rs7726159, P = 2.10 × 10−39; Region 3: rs2853677, P = 3.30 × 10−36 and PConditional = 2.36
× 10−8; Region 4: rs2736098, P = 3.87 × 10−12 and PConditional = 5.19 × 10−6, Region 5:
rs13172201, P = 0.041 and PConditional = 2.04 × 10−6; and Region 6: rs10069690, P = 7.49
× 10−15 and PConditional = 5.35 × 10−7) and one in the neighboring CLPTM1L gene (Region
2: rs451360; P = 1.90 × 10−18 and PConditional = 7.06 × 10−16). Between three and five
cancers mapped to each independent locus with both risk-enhancing and protective
effects. Allele-specific effects on DNA methylation were seen for a subset of risk loci,
indicating that methylation and subsequent effects on gene expression may contribute to
the biology of risk variants on 5p15.33. Our results provide strong support for extensive
pleiotropy across this region of 5p15.33, to an extent not previously observed in other
cancer susceptibility loci.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
12. Tilan JU, Krailo M, Barkauskas DA, Galli S, Mtaweh H, Long J, Wang H, Hawkins K, Lu C,
Jeha D, Izycka-Swieszewska E, Lawlor ER, Toretsky JA, Kitlinska JB. (2014) Systemic levels
of neuropeptide Y and dipeptidyl peptidase activity in patients with Ewing sarcoma –
Associations with tumor phenotype and survival. Cancer 121(5): 697–707.
http://dx.doi.org/10.1002/cncr.29090
Abstract
BACKGROUND
Ewing sarcoma (ES) is driven by fusion of the Ewing sarcoma breakpoint region 1 gene
(EWSR1) with an E26 transformation-specific (ETS) transcription factor (EWS-ETS), most
often the Friend leukemia integration 1 transcription factor (FLI1). Neuropeptide Y (NPY)
is an EWS-FLI1 transcriptional target; it is highly expressed in ES and exerts opposing
effects, ranging from ES cell death to angiogenesis and cancer stem cell propagation.
The functions of NPY are regulated by dipeptidyl peptidase IV (DPPIV), a hypoxiainducible enzyme that cleaves the peptide and activates its growth-promoting actions.
The objective of this study was to determine the clinically relevant functions of NPY by
identifying the associations between patients' ES phenotype and their NPY
concentrations and DPP activity.
METHODS
NPY concentrations and DPP activity were measured in serum samples from 223 patients
with localized ES and 9 patients with metastatic ES provided by the Children's Oncology
Group.
RESULTS
Serum NPY levels were elevated in ES patients compared with the levels in a healthy
control group and an osteosarcoma patient population, and the elevated levels were
independent of EWS-ETS translocation type. Significantly higher NPY concentrations
were detected in patients with ES who had tumors of pelvic and bone origin. A similar
trend was observed in patients with metastatic ES. There was no effect of NPY on
survival in patients with localized ES. DPP activity in sera from patients with ES did not
differ significantly from that in healthy controls and patients with osteosarcoma. However,
high DPP levels were associated with improved survival.
CONCLUSIONS
Systemic NPY levels are elevated in patients with ES, and these high levels are
associated with unfavorable disease features. DPPIV in serum samples from patients with
ES is derived from nontumor sources, and its high activity is correlated with improved
survival.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
13. Glover J, Krailo M, Tello T, Marina N, Janeway K, Barkauskas D, Fan TM, Gorlick R, Khanna
C, and the COG Osteosarcoma Biology Group. A Summary of the Osteosarcoma Banking
Efforts: A Report from the Children’s Oncology Group and the QuadW Foundation.
Pediatric Blood & Cancer 62(3): 450–455.
http://dx.doi.org/10.1002/pbc.25346
Abstract
Background
Survival rates of patients with osteosarcoma have remained stagnant over the last thirty
years. Better understanding of biology, new therapeutics, and improved biomarkers are
needed. The Children's Oncology Group (COG) addressed this need by developing one
of the largest osteosarcoma biorepositories ever, containing over 15,000 tumor and tissue
samples from over 1,500 patients.
Procedure
The biology study P9851 and the banking study AOST06B1 has enrolled 1,787 patients
(as of September, 2013). Clinical information was lacking on 510 patients on P9851, who
were not enrolled on a concurrent therapeutic trial. The value of these specimens was
diminished. The lack of statistical support available for biology projects slowed the
analysis of several critical studies. The QuadW Foundation, CureSearch, and the COG
formed the Childhood Sarcoma Biostatistics and Annotation Office (CSBAO) to provide
the infrastructure and address these needs by linking clinically annotated patient data to
archived tissue samples and to develop biostatistical support for childhood sarcoma
research.
Results
Originally 5.3% of samples from the 510 patients on P9851 not enrolled on a therapeutic
study had full clinical annotation. The efforts of the CSBAO have linked clinical annotation
to 90.8% of those specimens and provided statistical analyses to several studies that had
used COG samples. As a result, 24 biology studies in osteosarcoma have been
completed and published in peer-reviewed journals.
Conclusions
These samples and in-silico data are available to the research community for basic and
translational science projects to improve the biological understanding and treatment of
patients affected by osteosarcoma.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
14. Rosenblum JM, Wijetunga NA, Fazzari MJ, Krailo MK, Barkauskas DA, Gorlick R, Greally JM.
(2015) Predictive Properties of DNA Methylation Patterns in Primary Tumor Samples for
Osteosarcoma Relapse Status. Epigenetics 10(1): 31–39.
http://dx.doi.org/10.4161/15592294.2014.989084
Abstract
Osteosarcoma is the most common primary malignant bone tumor in children. Validated
biological markers for disease prognosis available at diagnosis are lacking. No genomewide DNA methylation studies linked to clinical outcomes have been reported in
osteosarcoma to the best of our knowledge. To address this, we tested the methylome at
over 1.1 million loci in 15 osteosarcoma biopsy samples obtained prior to the initiation of
therapy and correlated these molecular data with disease outcomes. At more than 17% of
the tested loci, samples obtained from patients who experienced disease relapse were
more methylated than those from patients who did not have recurrence while patients
who did not experience disease relapse had more DNA methylation at fewer than 1%. In
samples from patients who went on to have recurrent disease, increased DNA
methylation was found at gene bodies, intergenic regions and empirically-annotated
candidate enhancers, whereas candidate gene promoters were unusual for a more
balanced distribution of increased and decreased DNA methylation with 6.6% of gene
promoter loci being more methylated and 2% of promoter loci being less methylated in
patients with disease relapse. A locus at the TLR4 gene demonstrates one of strongest
associations between DNA methylation and 5 y event-free survival (P-value = 1.7 × 10−6),
with empirical annotation of this locus showing promoter characteristics. Our data indicate
that DNA methylation information has the potential to be predictive of outcome in pediatric
osteosarcoma, and that both promoters and non-promoter loci are potentially informative
in DNA methylation studies.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
15. Mirabello L, Yeager M, Mai PL, Gastier-Foster JM, Gorlick R, Khanna C, Patiño-Garcia A,
Sierrasesúmaga L, Lecanda F, Andrulis IL, Wunder JS, Gokgoz N, Barkauskas DA, Zhang X,
Vogt A, Jones K, Boland JF, Chanock SJ, Savage SA. Germline TP53 variants and
susceptibility to osteosarcoma. JNCI 107(7): djv101.
http://dx.doi.org/10.1093/jnci/djv101
Abstract
The etiologic contribution of germline genetic variation to sporadic osteosarcoma is not
well understood. Osteosarcoma is a sentinel cancer of Li-Fraumeni syndrome (LFS), in
which approximately 70% of families meeting the classic criteria have germline TP53
mutations. We sequenced TP53 exons in 765 osteosarcoma cases. Data were analyzed
with χ2 tests, logistic regression, and Cox proportional hazards regression models. We
observed a high frequency of young osteosarcoma cases (age <30 years) carrying a
known LFS- or likely LFS-associated mutation (3.8%) or rare exonic variant (5.7%) with
an overall frequency of 9.5%, compared with none in case patients age 30 years and
older (P < .001). This high TP53 mutation prevalence in young osteosarcoma cases is
statistically significantly greater than the previously reported prevalence of 3% (P =
.0024). We identified a novel association between a TP53 rare variant and metastasis at
diagnosis of osteosarcoma (rs1800372, odds ratio = 4.27, 95% confidence interval = 1.2
to 15.5, P = .026). Genetic susceptibility to young onset osteosarcoma is distinct from
older adult onset osteosarcoma, with a high frequency of LFS-associated and rare exonic
TP53 variants.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
16. Allen-Rhoades W, Kurenbekova L, Satterfield L, Parikh N, Fuja D, Shuck RL, Rainusso N,
Trucco M, Barkauskas DA, Jo E, Ahern C, Hilsenbeck S, Donehower LA, Yustein JT. Crossspecies identification of a plasma microRNA signature with applications for detection,
therapeutic monitoring, and prognosis in osteosarcoma. Cancer Medicine 4(7): 977–988.
http://dx.doi.org/10.1002/cam4.438
Abstract
Osteosarcoma (OS) is the primary bone tumor in children and young adults. Currently,
there are no reliable, noninvasive biologic markers to detect the presence or progression
of disease, assess therapy response, or provide upfront prognostic insights. MicroRNAs
(miRNAs) are evolutionarily conserved, stable, small noncoding RNA molecules that are
key posttranscriptional regulators and are ideal candidates for circulating biomarker
development due to their stability in plasma, ease of isolation, and the unique expressions
associated with specific disease states. Using a qPCR-based platform that analyzes more
than 750 miRNAs, we analyzed control and diseased-associated plasma from a
genetically engineered mouse model of OS to identify a profile of four plasma miRNAs.
Subsequent analysis of 40 human patient samples corroborated these results. We also
identified disease-specific endogenous reference plasma miRNAs for mouse and human
studies. Specifically, we observed plasma miR-205-5p was decreased 2.68-fold in mice
with OS compared to control mice, whereas, miR-214, and miR-335-5p were increased
2.37- and 2.69-fold, respectively. In human samples, the same profile was seen with miR205-5p decreased 1.75-fold in patients with OS, whereas miR-574-3p, miR-214, and miR335-5p were increased 3.16-, 8.31- and 2.52-fold, respectively, compared to healthy
controls. Furthermore, low plasma levels of miR-214 in metastatic patients at time of
diagnosis conveyed a significantly better overall survival. This is the first study to identify
plasma miRNAs that could be used to prospectively identify disease, potentially monitor
therapeutic efficacy and have prognostic implications for OS patients.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
17. Mirabello, et al. (2015) A genome-wide scan identifies variants in NFIB associated with
metastasis in patients with osteosarcoma. Cancer Discovery.
http://dx.doi.org/10.1158/2159-8290.CD-15-0125
Abstract
Metastasis is the leading cause of death in patients with osteosarcoma, the most common
pediatric bone malignancy. We conducted a multistage genome-wide association study of
osteosarcoma metastasis at diagnosis in 935 osteosarcoma patients to determine
whether germline genetic variation contributes to risk of metastasis. We identified an
SNP, rs7034162, in NFIB significantly associated with metastasis in European
osteosarcoma cases, as well as in cases of African and Brazilian ancestry (meta-analysis
of all cases: P = 1.2 × 10−9; OR, 2.43; 95% confidence interval, 1.83–3.24). The risk
allele was significantly associated with lowered NFIB expression, which led to increased
osteosarcoma cell migration, proliferation, and colony formation. In addition, a transposon
screen in mice identified a significant proportion of osteosarcomas harboring inactivating
insertions in Nfib and with lowered NFIB expression. These data suggest that germline
genetic variation at rs7034162 is important in osteosarcoma metastasis and that NFIB is
an osteosarcoma metastasis susceptibility gene.
SIGNIFICANCE: Metastasis at diagnosis in osteosarcoma is the leading cause of death in
these patients. Here we show data that are supportive for the NFIB locus as associated
with metastatic potential in osteosarcoma.
Osteosarcoma Abstracts
1. (Arndt, Carola et al (2012, May). Chemotherapy-associated Toxicities in Overweight and
Underweight Children with Osteosarcoma [abstract]. Poster session presented at the 25th
annual American Society of Pediatric Hematology Oncology meeting, New Orleans, LA.
2. (Glover, Jason et al (2013, May). The Children’s Oncology Group and QuadW Foundation
osteosarcoma banking experience [abstract]. Poster session presented at the annual
American Society of Clinical Oncology meeting, Chicago, IL.
Osteosarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
Ewing’s Sarcoma Publications
1. Borinstein SC, Barkauskas DA, Krailo M, Scher D, Scher L, Schlottmann S, Kallakury
B, Dickman PS, Pawel BR, West DC, Womer RB, Toretsky JA. Investigation of the insulinlike growth factor-1 signaling pathway in localized Ewing sarcoma: A report from the
Children’s Oncology Group. Cancer. 2011 Nov 1; 117(21): 4966–4976.
http://dx.doi.org/10.1002/cncr.26112
Abstract
BACKGROUND:
The insulin-like growth factor-1 (IGF-1) signaling pathway plays an important role in the
pathology of Ewing sarcoma (ES). Retrospective studies have suggested that levels of
IGF-1 and IGF binding protein 3 (IGFBP-3) are correlated with the outcome of patients
with ES.
METHODS:
The IGF-1 signaling pathway was investigated prospectively in 269 patients who had
localized, previously untreated ES. Serum samples were obtained at diagnosis, and
concentrations of IGF-1 and IGFBP-3 were determined by enzyme-linked immunosorbent
assays. In addition, immunohistochemistry (IHC) was performed to assay for
phosphorylated p70S6 kinase, protein kinase B (Akt), and forkhead box protein O1
(FOXO1) and to determine the presence of protein tyrosine phosphatase-L1 (PTPL1).
IHC findings along with IGF-1 and IGFBP-3 concentrations were correlated with age,
tumor location, sex, event-free survival, and overall survival.
RESULTS:
Patients aged >18 years tended to have higher levels of IGF-1 (P = .10), lower levels of
IGFBP-3 (P = .16), and decreased IGFBP-3:IGF-1 ratios (P = .01). No correlations were
observed between sex, tumor location, or outcomes and concentrations of IGF-1 or
IGFBP-3. Phosphorylation of p70S6 kinase, Akt, and FOXO1 was detected in the majority
of patient tissues but was not associated with age, sex, or tumor location. PTPL1 was
present in >80% of tumors and also was not correlated with age, sex, or tumor location.
There was no difference in survival with respect to the presence of phosphorylated p70S6
kinase, phosphorylated FOXO1, phosphorylated Akt, or PTPL1.
CONCLUSIONS:
The baseline IGFBP-3:IGF-1 ratio was correlated with age but did not affect the outcomes
of patients with ES. The authors concluded that additional investigation of the IGF-1
pathway is warranted in patients with ES, and especially in those who have received
treatment with IGF-1 receptor antibody inhibitors.
Ewing’s sarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
2. Borinstein SC, Beeler N, Block JJ, Gorlick R, Jedlicka P, Krailo M, Morris C, Phillips S, Siegal
G, Lawlor E, and Lessnick S. A decade in banking Ewing sarcoma: a report from the
Children’s Oncology Group. Frontiers in Oncology 3(57): 1, 2013 March 20.
http://dx.doi.org/10.3389/fonc.2013.00057
Abstract
Outcomes for patients with metastatic and recurrent Ewing sarcoma remain poor and a
better understanding of the biology of this malignancy is critical to the development of
prognostic biomarkers and novel therapies. Therefore, the Children’s Oncology Group
(COG) has created tissue banking protocols designed to collect high quality, clinically
annotated, tumor specimens that can be distributed to researchers to perform basic
science and correlative investigation. Data from the COG Ewing sarcoma tissue banking
protocols AEWS02B1 and its successor study AEWS07B1 were reviewed in this study.
Six-hundred and thirty five patients were enrolled on AEWS02B1 and 396 patients have
had tissue submitted to AEWS07B1. The average age of participation was 13.2 years.
About 86% were less than 19 years old and only 6% were greater than 21 years of age at
diagnosis. When compared to SEER data, approximately 18% of all cases and only 8% of
all patients >20 years old diagnosed with Ewing sarcoma annually in the United States
have had tumor banked. The majority of participants submitted formalin fixed, paraffin
embedded, primary tumor and blood samples. In total, fresh frozen tissue was submitted
for only 29% of cases. Only seven metastatic tumor samples have been collected.
Although the COG has been successful in collecting tumor samples from patients newly
diagnosed with Ewing sarcoma, fresh frozen tumor specimens from primary and
metastatic disease are critically needed, especially from young adult patients, in order to
conduct high quality basic science and translational research investigation with a goal of
developing better treatments.
Ewing’s sarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
3. Shukla NN, Schiffman JD, Reed DR, Davis IJ, Womer RB, Lessnick SL, Lawlor ER. Biomarkers
in Ewing sarcoma: the promise and challenge of personalized medicine. A report from the
Children’s Oncology Group. Frontiers in Oncology 3(141): 1–13, 2013 June 03.
http://dx.doi.org/10.3389/fonc.2013.00141
Abstract
A goal of the COG Ewing Sarcoma (ES) Biology Committee is enabling identification of
reliable biomarkers that can predict treatment response and outcome through the use of
prospectively collected tissues and correlative studies in concert with COG therapeutic
studies. In this report, we aim to provide a concise review of the most well-characterized
prognostic biomarkers in ES, and to provide recommendations concerning design and
implementation of future biomarker studies. Of particular interest and potentially high
clinical relevance are studies of cell-cycle proteins, sub-clinical disease, and copy number
alterations. We discuss findings of particular interest from recent biomarker studies and
examine factors important to the success of identifying and validating clinically relevant
biomarkers in ES. A number of promising biomarkers have demonstrated prognostic
significance in numerous retrospective studies and now need to be validated
prospectively in larger cohorts of equivalently treated patients. The eventual goal of
refining the discovery and use of clinically relevant biomarkers is the development of
patient specific ES therapeutic modalities.
Ewing’s sarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
4. Volchenboum SL, Andrade J, Huang L, Barkauskas DA, Krailo M, Womer RB, Ranft A,
Potratz J, Dirksen U, Triche TJ, Lawlor ER. (2015) Gene expression profiling of Ewing
sarcoma tumours reveals the prognostic importance of tumour-stromal interactions: a
report from the Children's Oncology Group. The Journal of Pathology: Clinical Research
1(2): 83–94 April, 2015.
http://dx.doi.org/10.1002/cjp2.9
Abstract
Relapse of Ewing sarcoma (ES) can occur months or years after initial remission, and
salvage therapy for relapsed disease is usually ineffective. Thus, there is great need to
develop biomarkers that can predict which patients are at risk for relapse so that therapy
and post-therapy evaluation can be adjusted accordingly. For this study, we performed
whole genome expression profiling on two independent cohorts of clinically annotated ES
tumours in an effort to identify and validate prognostic gene signatures. ES specimens
were obtained from the Children's Oncology Group and whole genome expression
profiling performed using Affymetrix Human Exon 1.0 ST arrays. Lists of differentially
expressed genes between survivors and non-survivors were used to identify prognostic
gene signatures. An independent cohort of tumours from the Euro-Ewing cooperative
group was similarly analysed as a validation cohort. Unsupervised clustering of gene
expression data failed to segregate tumours based on outcome. Supervised analysis of
survivors versus non-survivors revealed a small number of differentially expressed genes
and several statistically significant gene signatures. Gene-specific enrichment analysis
demonstrated that integrin and chemokine genes were associated with survival in
tumours where stromal contamination was present. Tumours that did not harbour stromal
contamination showed no association of any genes or pathways with clinical outcome.
Our results reflect the challenges of performing RNA-based assays on archived bone
tumour specimens. In addition, they reveal a key role for tumour stroma in determining ES
prognosis. Future biological and clinical investigations should focus on elucidating the
contribution of tumour:micro-environment interactions on ES progression and response to
therapy.
Ewing’s sarcoma publications
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QuadW-COG Childhood Sarcoma Biostatistics and Annotation Office
Publications / Abstracts
Soft Tissue Sarcoma Publications
1. Jacob AG, O'Brien D, Singh RK, Comiskey DF Jr, Littleton RM, Mohammad F, Gladman
JT, Widmann MC, Jeyaraj SC, Bolinger C, Anderson JR, Barkauskas DA, Boris-Lawrie
K, Chandler DS. Stress-induced isoforms of MDM2 and MDM4 correlate with high-grade
disease and an altered splicing network in pediatric rhabdomyosarcoma. Neoplasia 15(9):
1049–1063.
http://www.neoplasia.com/pdf/manuscript/v15i09/neo13286.pdf
Abstract
Pediatric rhabdomyosarcoma (RMS) is a morphologically and genetically heterogeneous
malignancy commonly classified into three histologic subtypes, namely, alveolar,
embryonal, and anaplastic. An issue that continues to challenge effective RMS patient
prognosis is the dearth of molecular markers predictive of disease stage irrespective of
tumor subtype. Our study involving a panel of 70 RMS tumors has identified specific
alternative splice variants of the oncogenes Murine Double Minute 2 (MDM2) and MDM4
as potential biomarkers for RMS. Our results have demonstrated the strong association of
genotoxic-stress
inducible
splice
forms
MDM2-ALT1
(91.6%
Intergroup
Rhabdomyosarcoma Study Group stage 4 tumors) and MDM4-ALT2 (90.9% MDM4ALT2-positive T2 stage tumors) with high-risk metastatic RMS. Moreover, MDM2-ALT1positive metastatic tumors belonged to both the alveolar (50%) and embryonal (41.6%)
subtypes, making this the first known molecular marker for high-grade metastatic disease
across the most common RMS subtypes. Furthermore, our results show that MDM2-ALT1
expression can function by directly contribute to metastatic behavior and promote the
invasion of RMS cells through a matrigel-coated membrane. Additionally, expression of
both MDM2-ALT1 and MDM4-ALT2 increased anchorage-independent cell-growth in soft
agar assays. Intriguingly, we observed a unique coordination in the splicing of MDM2ALT1 and MDM4-ALT2 in approximately 24% of tumor samples in a manner similar to
genotoxic stress response in cell lines. To further explore splicing network alterations with
possible relevance to RMS disease, we used an exon microarray approach to examine
stress-inducible splicing in an RMS cell line (Rh30) and observed striking parallels
between stress-responsive alternative splicing and constitutive splicing in RMS tumors.
Updated August 10, 2015
Soft tissue sarcoma publications
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