mSHOX2 Publications (please click on title for a short summary)

1. Dietrich et al., Performance evaluation of the DNA methylation biomarker SHOX2 for the aid in diagnosis of lung cancer based on the analysis of bronchial aspirates, Int J Oncol. 2012 Mar;40(3):825-32.
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The above study was performed in order to characterize the commercialized, CE marked Epi proLung BL Reflex Assay by validating the previously defined cut-off value for the mSHOX2 biomarker in bronchial lavage and by verifying the performance data of the test with respect to detection sensitivity and specificity. The patient cohort chosen for the study was independent than the cohort used in the previous feasibility study (Schmidt et al, 2010) and consisted of 125 confirmed lung cancer samples and 125 specific controls.

RESULTS

The results describe the test as a robust and reliable diagnostic tool for identifying patients with lung cancer using Saccomanno-fixed bronchial lavage specimens (AUC [95% confidence intervals] = 0.94 [0.91-0.98], sensitivity 78% [69-86]/specificity 96% [90-99]).

CONCLUSION

This test may be used as a diagnostic adjunct to existing clinical and pathological investigations in lung cancer.
2. Kneip C. et al., SHOX2 DNA Methylation is a Biomarker for the Diagnosis of Lung Cancer in Plasma, J Thorac Oncol. 2011;6
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This study aimed to develop a modified SHOX2 assay for use in a blood-based test and to analyze the performance of this optimized SHOX2 methylation assay in plasma. A training study (20 stage IV patients with lung cancer and 20 controls) was performed to show feasibility of detecting mSHOX2 in blood and to determine a methylation cutoff for patient classification. The resulting cutoff was verified in a testing study composed of 371 plasma samples from patients with lung cancer and controls.

Results

  • 112 out of 188 valid lung cancer patient samples were SHOX2 positive, resulting in an overall sensitivity of 60 %.
  • 16 out of 155 valid controls were SHOX2 methylation positive, resulting in a specificity of 90 %.
  • SCLC (80 %) and squamous cell carcinoma (63 %) were detected at the highest sensitivity as compared to adenocarcinomas.

Conclusion

SHOX2 DNA methylation is a biomarker for detecting the presence of malignant lung disease in blood plasma form patients with lung cancer. 
3. Field J. et al., SHOX2 DNA Methylation is a Biomarker for the Diagnosis of Lung Cancer in Plasma, 102 Annual Meeting AACR, 2011, Abstract #4158/20
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This study aimed to analyze the performance of the SHOX2 methylation biomarker in plasma. In the study 371 patients were enrolled, 155 patients with benign lung disease and 188 lung cancer patients.

Results

  • 116 out of 188 cases were SHOX2 positive, resulting in an overall sensitivity of 62 %.
  • 16 out of 155 controls were SHOX2 methylation positive, resulting in a specificity of 90 %.
  • SCLC (80 %) and squamous cell carcinoma (63 %) were detected at the highest sensitivity as compared to adenocarcinomas.

Conclusion

SHOX2 DNA methylation is a sensitive and specific biomarker for detecting the presence of malignant lung disease in blood plasma, in particular SCLC and
squamous cell carcinoma. A blood-based test for SHOX2 methylation could be a useful tool for confirming malignant disease in patients suspected of having
lung cancer.
4. Schneider K. U. et al., Correlation of  SHOX2 Gene Amplification and DNA Methylation in Lung Cancer Tumors. BMC Cancer 2011, 11 (1):102st
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This study aimed to investigate if the hypermethylation of SHOX2 in lung cancer patients correlates with SHOX2 gene expression and/or copy number alterations. The study was comprised of matched morphologically normal lung tissue and tumor tissue from 55 lung cancer patients who underwent surgery. Surgical samples were obtained from the ELK Berlin Chest Hospital, Germany. 

Results

Hypermethylation of the SHOX2 locus in tumor tissue as compared to the matched NAT from the same patient was detected in 96 % of tumors from the 55 lung cancer patients. This correlated highly significantly with the frequent occurrence of copy number amplification (p < 0.0001), while expression of the SHOX2 gene showed no difference

Conclusion

Frequent gene amplification correlated with hypermethylation of the SHOX2 gene locus. This concerted effect qualifies SHOX2 DNA methlyation as a biomarker for lung cancer diagnosis, especially when sensitive detection is needed, e. g. in bronchial lavage or blood samples.
5. Schmidt B. et al., SHOX2 DNA Methylation is a Biomarker for the Diagnosis of Lung Cancer Based on Bronchial Aspirates, BMC Cancer 2010, 10: 600
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This study aimed to show that SHOX2 DNA methylation is a tumor marker in patients with suspected lung cancer by using bronchial fluid aspirated during bronchoscopy. In the study 523 patients were enrolled, 242 patients with benign lung disease and 281 lung cancer patients. Fresh-frozen and saccomanno-fixed samples were used to show the tumor marker performance in different sample types of clinical relevance. Samples were obtained from Charité University Hospital in Berlin, Germany and from the Roy Castle Lung Cancer Research Program, Liverpool, UK.

Results

Valid measurements were obtained from a total of 523 patient samples (242 controls, 281 cases). Determination of SHOX2 DNA methylation levels allowed us to distinguish between malignant and benign lung disease, i. e. abscesses, infections, obstructive lung diseases, sarcoidosis, scleroderma, stenoses, at high specificity (68 % sensitivity [95 % CI 62–73 %], 95 % specificity [95 % CI 91–97 %]). 

Conclusions

Hypermethylation of SHOX2 in bronchial aspirates appears to be a clinically useful tumor marker for identifying subjects with lung carcinoma, especially if histological and cytological findings after bronchoscopy are ambiguous.
6. Field J. et al., SHOX2, a Novel Biomarker for the Detection of Lung Cancer in Bronchial Fluid, 100th Annual Meeting AACR, 2009, Abstract #254
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This retrospective clinical study was designed to demonstrate that a recently developed HeavyMethyl (HM) sensitive detection methylation assay for the SHOX2 gene could be translated into clinical practice as a marker to diagnose bronchial carcinoma in patients undergoing workup for suspected lung cancer. Methylation of the SHOX2 gene was measured in two independent populations of patients undergoing workup for suspected lung cancer. Cases had histologically confirmed lung cancer while controls had no malignant disease at that time and at least one year after. One population consisted of saccomanno-fixed bronchial lavage samples. The other population contained fresh-frozen lavage samples from patients that showed inconclusive or cytology negative results after first bronchoscopy. 

Results

Sufficient DNA could be extracted from 423 samples (151 fresh-frozen; 272 saccomanno-fixed). SHOX2 was hypermethylated in cancer patients as compared to the control group for both populations. Using a predefined cut off, the sensitivity for the saccomanno-fixed and the fresh-frozen lavage samples with inconclusive cytology results was determined to be 62 % and 53 %, respectively. The specificity was 99 % for both populations. Squamous carcinoma had a particularly high detection rate with 75 % (63/84) found to be SHOX2 methylation positive at a specificity of 99 % (2/186). 

Conclusions

Saccomanno-fixed and fresh-frozen lavage material is suitable for analysis, making the workflow suitable for clinical use. The detection of cytology negative cases suggests that DNA methylation of SHOX2 contains valuable clinical information.