Characterization of Three Novel H3F3A-mutated Giant Cell Tumor Cell Lines and Targeting of Their Wee1 Pathway.

The giant cell tumor of bone (GCTB) is a locally aggressive primary bone tumor that is composed of mononuclear stroma cells, scattered macrophages, and multinucleated osteoclast-like giant cells which cause pathologic osteolysis. The stroma cells represent the neoplastic population of the tumor and are characterized by the H3F3A mutation G34W. This point mutation is regarded as the driver mutation of GCTB. We have established three new stable H3F3A mutated GCTB cell lines: U-GCT1, U-GCT2, and U-GCT3M. MK-1775 is a Wee1-kinase inhibitor which has been used for blocking of sarcoma growth. In the cell lines we detected Wee1, Cdk1, Cyclin B1, H3K36me3, and Rrm2 as members of the Wee1 pathway. We analyzed the effect of MK-1775 and gemcitabine, alone and in combination, on the growth of the cell lines. The cell lines showed a significant reduction in cell proliferation when treated with MK-1775 or gemcitabine. The combination of both agents led to a further significant reduction in cell proliferation compared to the single agents. Immunohistochemical analysis of 13 GCTB samples revealed that Wee1 and downstream-relevant members are present in GCTB tissue samples. Overall, our work offers valuable new tools for GCTB studies and presents a description of novel biomarkers and molecular targeting strategies.

H3F3A mutation in giant cell tumour of the bone is detected by immunohistochemistry using a monoclonal antibody against the G34W mutated site of the histone H3.3 variant.

AIMS: Giant cell tumour of the bone (GCTB) is a neoplasm predominantly of long bones characterized by the H3F3A mutation G34W. Conventional diagnosis is challenged by the tumour's giant cell-rich morphology, which overlaps with other giant cell-containing lesions of the bone. Recently, a monoclonal antibody specific for the H3F3A mutation has been generated. Our aim was to test this antibody on a cohort of giant cell-containing lesions. METHODS AND RESULTS: We used the antibody for analysis of 22 H3F3A-mutated GCTB, including two patients with recurrences; for comparison we analysed a cohort of 36 H3F3A wild-type giant cell-rich lesions of the bone and soft tissue, containing one brown tumour, six aneurysmal bone cysts (ABC), six chondroblastomas, five non-ossifying-fibromas, two fibrous dysplasias, nine tenosynovial giant cell tumours, one giant cell-rich sarcoma and six osteosarcomas. Furthermore, among the 22 mutated cases, we included one GCTB with two recurrences and lung metastases; the patient was treated with the anti-receptor activator of nuclear factor κB (RANK) ligand denosumab. We show that all 22 H3F3A-mutated GCTB display strong nuclear H3.3 G34W staining in the neoplastic component, while the osteoclastic giant cells are negative. 36 H3F3A wild-type lesions are negative. The GCTB treated with denosumab revealed a reduction in the H3.3 G34W-positive tumour cells and a decrease in osteoclastic giant cells accompanied by matrix and osteoid formation. CONCLUSIONS: We conclude that positive H3.3 G34W staining is a specific and sensitive method for detection of H3F3A-mutated GCTB. Denosumab treatment leads to a pathomorphosis of the lesion characterized by matrix and osteoid producing H3.3 G34W-negative stromal cells.

Wireless substitution: state-level estimates from the National Health Interview Survey, 2012.

OBJECTIVES: This report updates subnational estimates of the percentage of adults and children living in households that do not have a landline telephone but have at least one wireless telephone (i.e., wireless-only households). State-level estimates for 2012 are presented, along with estimates for selected U.S. counties and groups of counties, for other household telephone service use categories (e.g., those that had only landlines and those that had landlines yet received all or almost all calls on wireless telephones), and for one earlier 12-month period (July 2011-June 2012). METHODS: Small-area statistical modeling techniques were used to estimate the prevalence of adults and children living in households with various household telephone service types for 93 disjoint geographic areas that make up the United States. This modeling was based on 2007-2012 data from the National Health Interview Survey, 2006-2011 data from the American Community Survey, and auxiliary information on the number of listed telephone lines per capita in 2007-2012. RESULTS: The prevalence of wireless-only adults and children varied substantially across states. State-level estimates for 2012 ranged from 19.4% (New Jersey) to 52.3% (Idaho) of adults and from 20.6% (New Jersey) to 63.4% (Mississippi) of children.

Wireless substitution: state-level estimates from the National Health Interview Survey, 2010-2011.

OBJECTIVES: This report updates subnational estimates of the percentage of adults and children living in households without a landline telephone but with at least one wireless telephone (i.e., wireless-only households). State-level estimates for 2011 are presented, as well as estimates for selected U.S. counties and groups of counties, for other household telephone service use categories (e.g., those that had only landlines and those that had landlines yet received all or almost all calls on wireless telephones), and for two earlier 12-month periods (January-December 2010 and July 2010-June 2011). METHODS: Small-area statistical modeling techniques were used to estimate the prevalence of adults and children living in households with various household telephone service types for 93 disjoint geographic areas that make up the United States. This modeling was based on 2007-2011 data from the National Health Interview Survey, 2006-2010 data from the American Community Survey, and auxiliary information on the number of listed telephone lines per capita in 2007-2011. RESULTS: The prevalence of wireless-only adults and children varied substantially across states. State-level estimates for 2011 ranged from 15.3% (Rhode Island) to 44.6% (Idaho) of adults and from 15.2% (Rhode Island) to 58.6% (Mississippi) of children.

Wireless substitution: state-level estimates from the National Health Interview Survey, January 2007-June 2010.

OBJECTIVES: This report presents state-level estimates of the percentage of adults and children living in households that did not have a landline telephone but did have at least one wireless telephone. National estimates for the 12-month time period from July 2009 through June 2010 indicate that 23.9% of adults and 27.5% of children were living in these wireless-only households. Estimates are also presented for selected U.S. counties and groups of counties, for other household telephone service use categories (e.g., those that had only landlines and those that had landlines yet received all or almost all calls on wireless telephones), and for 12-month time periods since January-December 2007. METHODS: Small-area statistical modeling techniques were used to estimate the prevalence of adults and children living in households with various household telephone service types for 93 disjoint geographic areas that make up the entire United States. This modeling was based on January 2007-June 2010 data from the National Health Interview Survey, 2006-2009 data from the American Community Survey, and auxiliary information on the number of listed telephone lines per capita in 2007-2010. RESULTS: The prevalence of wireless-only adults and children varied substantially across states. State-level estimates for July 2009-June 2010 ranged from 12.8% (Rhode Island and New Jersey) to 35.2% (Arkansas) of adults and from 12.6% (Connecticut and New Jersey) to 46.2% (Arkansas) of children. For adults, the magnitude of the increase from 2007 to 2010 was lowest in New Jersey (7.2 percentage points) and highest in Arkansas (14.5 percentage points).

Reevaluating the need for concern regarding noncoverage bias in landline surveys.

OBJECTIVES: We used recent data to reexamine whether the exclusion of adults from households with no telephone or only wireless phones may bias estimates derived from health-related telephone surveys. METHODS: We calculated the difference between estimates for the full population of adults and estimates for adults with landline phones; data were from the 2007 National Health Interview Survey. RESULTS: When data from landline telephone surveys were weighted to match demographic characteristics of the full population, bias was generally less than 2 percentage points (range = 0.1-2.4). However, among young adults and low-income adults, we found greater bias (range = 1.7-5.9) for estimates of health insurance, smoking, binge drinking, influenza vaccination, and having a usual place for care. CONCLUSIONS: From 2004 to 2007, the potential for noncoverage bias increased. Bias can be reduced through weighting adjustments. Therefore, telephone surveys limited to landline households may still be appropriate for health surveys of all adults and for surveys of subpopulations regarding health status. However, for some behavioral risk factors and health care service use indicators, caution is warranted when using landline surveys to draw inferences about young or low-income adults.

Wireless substitution: state-level estimates from the National Health Interview Survey, January-December 2007.

OBJECTIVES: This report presents state-level estimates of the percentage of households that do not have a landline telephone, but do have at least one wireless telephone. These wireless-only households made up 14.7% of U.S. households in 2007. The report also presents state-level estimates of the percentage of adults living in wireless-only households. These wireless-only adults made up 13.6% of U.S. adults in 2007. METHODS: A two-sample modeling strategy was used to estimate the prevalence of wireless-only households and adults by state. This modeling was based on data from the 2007 National Health Interview Survey and the 2008 Current Population Survey's Annual and Social Economic Supplement. RESULTS: The results show that the prevalence of wireless-only households and adults in 2007 varied substantially across states. State-level estimates ranged from 5.1% (Vermont) to 26.2% (Oklahoma) of households and from 4.0% (Delaware) to 25.1% (Oklahoma) of adults. In addition, approximately one out of four adults (25.4%) living in the District of Columbia were wireless-only.

Telephone coverage and health survey estimates: evaluating the need for concern about wireless substitution.

OBJECTIVES: We sought to determine whether the exclusion of adults without landline telephones may bias estimates derived from health-related telephone surveys. METHODS: We took data from the 2004 and 2005 National Health Interview Survey and used logistic regression to compare the odds of behavioral risk factors and health care service use for adults with landline telephones to those for adults with only wireless telephones and adults without any telephone service. RESULTS: When interviewed, 7.2% of adults, including those who did and did not have wireless telephones, did not have landline telephones. Relative to adults with landline telephones, adults without landline telephones had greater odds of smoking and being uninsured, and they had lower odds of having diabetes, having a usual place for medical care, and having received an influenza vaccination in the past year. CONCLUSIONS: As people substitute wireless telephones for landline telephones, the percentage of adults without landline telephones has increased significantly but is still low, which minimizes the bias resulting from their exclusion from telephone surveys. Bias greater than 1 percentage point is expected only for estimates of health insurance, smoking, binge drinking, having a usual place for care, and receiving an influenza vaccination.

Estimating the prevalence of uninsured children: an evaluation of data from the National Survey of Children with Special Health Care Needs, 2001. Data evaluation and methods research.

UNLABELLED: The National Survey of Children with Special Health Care Needs revealed that 8.3% of children under 18 years of age were uninsured, a rate lower than the rate estimated by other national surveys. This report presents the results of an evaluation of the quality of this estimate, based on analyses of non-response, question design, interviewer and respondent effects, and the weighing and estimation process. National and State-level statistics on health insurance coverage for children with special health care needs (CSHCN) and for children without special needs are included in an appendix. SOURCE OF DATA: The National Survey of CSHCN is a survey module of the State and Local Area Integrated Telephone Survey. This survey of parents and guardians collected health insurance coverage information for a national sample of 215, 162 children. Data were collected from October 2000 through April 2002. RESULTS: Compared with other surveys, weighted data from the National Survey of CSHCN describe a population with a slightly larger proportion of Hispanic children and children from households with higher incomes. The National Survey of CSHCN was also the only survey to use a child-level design: A randomized experiment that varied the health insurance questions found that repeating the coverage questions for each child produced lower unisurance rates than household-level questions that first asked if anyone in the househol was insured. CONCLUSION: Question design differences explain much of the discrepancy between survey estimates of the uninsurance rate, but a definitive conclusion regarding the relative accuracy of the uninsurance rates is not possible.