Δευτέρα 24 Οκτωβρίου 2016

Touch imprint cytology with massively parallel sequencing (TIC-seq): a simple and rapid method to snapshot genetic alterations in tumors

Abstract

Identifying genetic alterations in tumors is critical for molecular targeting of therapy. In the clinical setting, formalin-fixed paraffin-embedded (FFPE) tissue is usually employed for genetic analysis. However, DNA extracted from FFPE tissue is often not suitable for analysis because of its low levels and poor quality. Additionally, FFPE sample preparation is time-consuming. To provide early treatment for cancer patients, a more rapid and robust method is required for precision medicine. We present a simple method for genetic analysis, called touch imprint cytology combined with massively paralleled sequencing (touch imprint cytology [TIC]-seq), to detect somatic mutations in tumors. We prepared FFPE tissues and TIC specimens from tumors in nine lung cancer patients and one patient with breast cancer. We found that the quality and quantity of TIC DNA was higher than that of FFPE DNA, which requires microdissection to enrich DNA from target tissues. Targeted sequencing using a next-generation sequencer obtained sufficient sequence data using TIC DNA. Most (92%) somatic mutations in lung primary tumors were found to be consistent between TIC and FFPE DNA. We also applied TIC DNA to primary and metastatic tumor tissues to analyze tumor heterogeneity in a breast cancer patient, and showed that common and distinct mutations among primary and metastatic sites could be classified into two distinct histological subtypes. TIC-seq is an alternative and feasible method to analyze genomic alterations in tumors by simply touching the cut surface of specimens to slides.

Thumbnail image of graphical abstract

In this study, we evaluated touch imprint cytology with massively parallel sequencing (touch imprint cytology [TIC]-seq) as an improved method for the genetic analysis of cancer patients. Our findings provide clear evidence that TIC-seq is a more robust and efficient method than FFPE-derived DNA analysis for the detection of somatic mutations in cancer, and capture intratumoral heterogeneity among primary and metastatic sites. TIC-seq has potential application for small-volume samples obtained by a number of standard clinical approaches, thus enabling more rapid diagnosis and treatment.



from Cancer via ola Kala on Inoreader http://ift.tt/2eJLgSL
via IFTTT

Δεν υπάρχουν σχόλια:

Δημοσίευση σχολίου