Using In Situ Padlock Probe Technology to Detect mRNA Splice Variants in Tumor Cells

2020 ◽  
pp. 361-378
Author(s):  
Lilli Hofmann ◽  
Thomas Kroneis ◽  
Amin El-Heliebi
Keyword(s):  
Tumor Cells ◽  
Splice Variants ◽  
Padlock Probe ◽  
Mrna Splice

scholarly journals In Situ Detection and Quantification of AR-V7, AR-FL, PSA, and KRAS Point Mutations in Circulating Tumor Cells

2018 ◽  
Vol 64 (3) ◽  
pp. 536-546 ◽  
Author(s):  
Amin El-Heliebi ◽  
Claudia Hille ◽  
Navya Laxman ◽  
Jessica Svedlund ◽  
Christoph Haudum ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Specific Antigen ◽  
Padlock Probe ◽  
Castration Resistant Prostate Cancer ◽  
Kras Mutations ◽  
Padlock Probes ◽  
Kras Status

Abstract BACKGROUND Liquid biopsies can be used in castration-resistant prostate cancer (CRPC) to detect androgen receptor splice variant 7 (AR-V7), a splicing product of the androgen receptor. Patients with AR-V7-positive circulating tumor cells (CTCs) have greater benefit of taxane chemotherapy compared with novel hormonal therapies, indicating a treatment-selection biomarker. Likewise, in those with pancreatic cancer (PaCa), KRAS mutations act as prognostic biomarkers. Thus, there is an urgent need for technology investigating the expression and mutation status of CTCs. Here, we report an approach that adds AR-V7 or KRAS status to CTC enumeration, compatible with multiple CTC-isolation platforms. METHODS We studied 3 independent CTC-isolation devices (CellCollector, Parsortix, CellSearch) for the evaluation of AR-V7 or KRAS status of CTCs with in situ padlock probe technology. Padlock probes allow highly specific detection and visualization of transcripts on a cellular level. We applied padlock probes for detecting AR-V7, androgen receptor full length (AR-FL), and prostate-specific antigen (PSA) in CRPC and KRAS wild-type (wt) and mutant (mut) transcripts in PaCa in CTCs from 46 patients. RESULTS In situ analysis showed that 71% (22 of 31) of CRPC patients had detectable AR-V7 expression ranging from low to high expression [1–76 rolling circle products (RCPs)/CTC]. In PaCa patients, 40% (6 of 15) had KRAS mut expressing CTCs with 1 to 8 RCPs/CTC. In situ padlock probe analysis revealed CTCs with no detectable cytokeratin expression but positivity for AR-V7 or KRAS mut transcripts. CONCLUSIONS Padlock probe technology enables quantification of AR-V7, AR-FL, PSA, and KRAS mut/wt transcripts in CTCs. The technology is easily applicable in routine laboratories and compatible with multiple CTC-isolation devices.

Abstract 2029: Wild type BRCA2 mRNA splice variants in normal and breast tumor cells

2011 ◽  
Author(s):  
Toshio F. Yoshimatsu ◽  
James D. Fackenthal ◽  
Olufunmilayo I. Olopade
Keyword(s):  
Tumor Cells ◽  
Breast Tumor ◽  
Splice Variants ◽  
Wild Type ◽  
Breast Tumor Cells ◽  
Mrna Splice

scholarly journals Insulin-inducible changes in insulin receptor mRNA splice variants.

1994 ◽  
Vol 269 (49) ◽  
pp. 30769-30772
Author(s):  
S M Sell ◽  
D Reese ◽  
V M Ossowski
Keyword(s):  
Insulin Receptor ◽  
Splice Variants ◽  
Mrna Splice ◽  
Receptor Mrna

scholarly journals Bdnf mRNA splice variants differentially impact CA1 and CA3 dendrite complexity and spine morphology in the hippocampus

2017 ◽  
Vol 222 (7) ◽  
pp. 3295-3307 ◽  
Author(s):  
Kristen R. Maynard ◽  
John W. Hobbs ◽  
Mahima Sukumar ◽  
Alisha S. Kardian ◽  
Dennisse V. Jimenez ◽  
...  
Keyword(s):  
Splice Variants ◽  
Bdnf Mrna ◽  
Spine Morphology ◽  
Mrna Splice

The expression of TP53 mRNA splice variants relates to progression of cancer in human colorectal cancer

Pathology ◽  
2014 ◽  
Vol 46 ◽  
pp. S117
Author(s):  
Hiroyuki Nozaka ◽  
Kentaro Endo ◽  
Akifumi Mayama ◽  
Kodai Takahashi ◽  
Hideki Takami ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Splice Variants ◽  
Human Colorectal Cancer ◽  
Mrna Splice
Sign in / Sign up

Export Citation Format

Share Document