Moffitt Cancer Center Morsani Molecular Lab Researchers Create New Laboratory Test to Better Diagnose Metastatic Bladder Cancer

Moffitt researchers reported the discovery of a new 19 gene expression signature that appears to have value in improving the accuracy of urothelial cancer diagnosis at the USCAP 2018 meeting in Vancouver.

The correct diagnosis of urothelial cancer, especially when metastatic can be very challenging for the surgical pathologist Urothelial carcinoma (UC) can mimic a poorly squamous cell carcinoma (SCC) and it may be difficult to distinguish the two especially in metastatic sites.

Metastatic cancer imaged with PET scan

 

It is important to distinguish the two cancers for proper patient management. These two cancers have an overlapping immunohistochemical profile (both positive for CK5/6, CK7, p63 and p40). GATA3 although relatively specific for UC, can also be expressed in SCC.

It is important to distinguish urothelial carcinoma from other epithelial cancers to ensure proper diagnosis and patient managment

Gene expression analysis is a powerful method that can measure the expression level of thousands of mRNA messages simultaneously in a tissue biopsy specimen. Analysis of data from this technique, when coupled with bioinformatics analysis can produce powerful classification algorithms which could be used in a CLIA laboratory to improve the accuracy of current diagnostic approaches.

Affymetrix chips for rapid gene expression

The Moffitt researchers analyzed 161 UC, 38 head and neck cases, and 268 lung cancer SCC cases from Moffitts Total Cancer Care (TCC) database to create the initial algorithm. The data was collected on the a specially modified HuRSTA chip containing 60607 probe sets covering 26356 genes. An algorithm was developed from the 19 top most differentially expressed genes using principal component analysis (PCA). The PCA model performed at above 98% sensitivity and specificity correctly identifying the histological origin of the specimens on self validation.

PCA analysis separating tumor types using 19 gene signature

The signature was further validated on publicly available external datasets at GEO datasets, with 96 cases of H&N SCC from GSE31056, 18 lung SCCs from GSE10245 and 93 UC from GSE31684, total of 207 cases.This signature correctly identified 112 of the 114 publicly available SCCs with gene data (96 H&N and 16 lung) as SCC and all of 93 publicly available UCs with gene data as UC

The identification of a gene classification algorithm will enable improved classification of these challenging lesions, particularly in the metastatic setting.

The Moffitt investigators will continue to evaluate this new signature and propose to develop it into a routine diagnostic assay. In addition, deeper understanding of the molecular basis of urothelial cancer and how it differs at the molecular level potentially opens new approaches to therapy.

An assay like this could be far superior than classical IHC as the complexity of gene expression will give multiple insights into not only tissue of orign but also other aspects of the cancer biology such as immune status.

Understanding the molecular basis of urothelial cancer will provide insights into novel treatment opportunity

The work was supported by the Moffitt Morsani Molecular Laboratories

Presented at the 2018 USCAP meeting in Vancouver Canada

Gene Biomarker Signature for Distinguishing Urothelial Carcinoma from Squamous Cell Carcinoma. Jasreman Dhillon, MD Yin Xiong, Ph.D. Anthony Magliocco, MD, Soner Altiok, MD, PhD, H. Lee Moffitt Cancer Center, Tampa FL

MOFFITT NGS STAR* Enters Clinical Service

Moffitt’s latest NGS sequencing assay the Moffitt STAR (Solid Tumor Actionable Result) panel was validated by the Moffitt Morsani Molecular Laboratory and launched into service this month at the busy Florida Comprehensive Cancer Center in Tampa.

The assay is based on Illumina’s TruSight Tumor 170 assay which is a next-generation sequencing assay designed to cover 170 genes that are commonly designated as drivers in solid tumors. The assay evaluates both DNA and RNA and focuses on detecting actionable mutations which include SNV, dels, insertions, amplifications, and translocations. Such alterations are the target for many new targetable therapies including anti-EGFR agents, anti BRAF therapies and treatments targeting the Tropomyosin Receptor Kinase fusions (TRK) such as Larotrectinib.

Many key actionable mutations only occur rarely, making detection by single marker tests problematic and wasteful. However, the Moffitt STAR assay now allows the Moffitt molecular laboratory to screen patient tumors for multiple targetable mutations efficiently in a single test using a relatively small amount of nucleic acid extracted from routine formalin fixed, paraffin embedded tissues (FFPE). This important advance enables the Moffitt molecular diagnostic laboratory to effectively evaluate a patient for eligibility to receive treatment with a FDA approved targeted therapy, or be considered for clinical trial enrollment. Moffitt STAR is essentially an “All in one” test that can provide multiple functions.

Moffitt NGS STAR* is an exciting new “all in one” technology advance for Moffitt Cancer Center patients enabling rapid assessment of their tumors for presence of key mutations directing selection of effective approved targeted therapies or for qualification to enroll in the latest generation of clinical trials

Evidence is also emerging the assay, despite its mid size, Moffitt STAR could also reliably measure tumor mutational load and microsatellite instability. These molecular features are often associated with potential response to the latest immune check point inhibitors such as Pembrolizumab which has recently received FDA approval for use in tumors with high microsatellite instability.

Moffitt NGS STAR also provides information on tumor mutational burden and microsatellite instability- key features which may drive patient response to the latest immuno-oncology check point inhibitor therapies

Moffitt NGS STAR can also detect mutations in BRCA genes, a molecular feature that may predict response to parp inhibitors such as olaparib.

Moffitt NGS STAR can be performed on as little as 40ng of input nucleic acid.

Development and launch of Moffitt NGS STAR was made possible through collaboration with industry partners PierianDx and Illumina Inc.

The Moffitt Cancer center is one of the largest in the United States, is consistently ranked in the top cancer centers by U.S. News & World Report. Moffitt Cancer Center has a mission to “contribute to the prevention and cure of cancer” and the vision ” to transform cancer care through service, science, and partnership”

For further details contact anthony.magliocco@moffitt.org

TRIPLE NEGATIVE BREAST CANCER IS OVER DIAGNOSED

By Dr. Anthony Magliocco

Getting a second opinion for a cancer diagnosis is highly recommended, but even more so if you face triple negative breast cancer, which can be aggressive and difficult to treat.

A new study led by Moffitt Cancer Center pathologist Dr. Marilin Rosa shows that triple negative breast cancer may be frequently overdiagnosed and reclassified after expert review and biomarker retesting. Moffitt investigators presented the data at the 2018 United States & Canadian Academy of Pathology Annual Meeting in Vancouver.

Researchers reviewed over 560 cases of breast cancer referred to Moffitt and found that 113 were initially classified as triple negative by external evaluation. After biomarker retesting, about 28 percent of the triple negative cases were reclassified as hormone receptor positive.

Moffitt’s study demonstrates the value of biomarker retesting for triple negative breast cancers before selecting an appropriate treatment plan. A second opinion that changes your diagnosis can have a huge impact on survival.

In triple negative breast cancer, the three most common types of receptors known to fuel most breast cancer growth — estrogen, progesterone and the HER-2 gene — are not present. This makes common treatments such as hormone therapy and drugs that target the three missing receptors ineffective.

Up to 20 percent of breast cancer diagnoses are triple negative and are more likely to affect younger patients, blacks, Hispanics and those with a BRCA1 gene mutation. This disease is also more likely to spread and recur.

The takeaway: Having an accurate cancer diagnosis is critical to planning appropriate treatment. If you are diagnosed with triple negative breast cancer, consider getting a second opinion before starting a treatment plan.