Molecular signatures of mu opioid receptor and somatostatin receptor 2 in pancreatic cancer

Jorand, Raphaël; Biswas, Sunetra; Wakefield, Devin L.; Tobin, Steven J.; Golfetto, Ottavia; Hilton, Kelsey; Ko, Michelle; Ramos, Joe W.; Small, Alexander R.; Singh, Gagandeep; Jovanovic-Talisman, Tijana

doi:10.1091/mbc.e16-06-0427

Cited by 31 publications

(27 citation statements)

References 97 publications

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“…Coverslips (25-mm diameter, #1.5; Warner Instruments, Hamden, CT) were cleaned as described before 30 . Clean coverslips were stored in sterile 35-mm tissue culture dishes for touch preparation or cell culture.…”

Section: Methodsmentioning

confidence: 99%

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Single molecule localization microscopy coupled with touch preparation for the quantification of trastuzumab-bound HER2

Tobin

Wakefield

Jones

et al. 2018

Sci Rep

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All breast cancers are assessed for levels of human epidermal growth factor receptor 2 (HER2). Fluorescence in situ hybridization (FISH) and immunohistochemistry are currently used to determine if a patient is eligible for anti-HER2 therapy. Limitations of both tests include variability and relatively long processing times. Additionally, neither test determines whether HER2 contains the extracellular domain. While truncated in some tumors, this domain is required for binding of the therapeutic antibody trastuzumab. Here, trastuzumab was used to directly detect HER2 with quantitative single molecule localization microscopy (qSMLM). In proof of concept studies, our new method rapidly quantified both HER2 density and features of nano-organization. In cultured cells, the method was sensitive to subtle variations in HER2 expression. To assess patient samples, we combined qSMLM with tissue touch preparation (touch prep-qSMLM) and examined large areas of intact membranes. For cell lines and patient samples, HER2 copy numbers from FISH showed a significant positive correlation with detected densities from qSMLM and trended with HER2 cluster occupancy.

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Section: Methodsmentioning

confidence: 99%

“…These techniques are also compatible with tissue specimen imaging. SMLM was used to detect HER2 in paraffin embedded tissue sections 29 and to determine how G-protein coupled receptors organize in fresh-frozen human tumor samples 30 .…”

Section: Introductionmentioning

confidence: 99%

Single molecule localization microscopy coupled with touch preparation for the quantification of trastuzumab-bound HER2

Tobin

Wakefield

Jones

et al. 2018

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…25 mm #1.5 coverslips (Warner Instruments, Hamden, CT) were cleaned as previously described. 60 As shown in Scheme S1 , His-tagged proteins were covalently attached to diazotized surfaces. Surfaces were first incubated with concentrated HCl for 2 min, followed by several rinses with distilled water and absolute ethanol.…”

Section: Methodsmentioning

confidence: 99%

A Platform To Enhance Quantitative Single Molecule Localization Microscopy

et al. 2018

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Quantitative single molecule localization microscopy (qSMLM) is a powerful approach to study in situ protein organization. However, uncertainty regarding the photophysical properties of fluorescent reporters can bias the interpretation of detected localizations and subsequent quantification. Furthermore, strategies to efficiently detect endogenous proteins are often constrained by label heterogeneity and reporter size. Here, a new surface assay for molecular isolation (SAMI) was developed for qSMLM and used to characterize photophysical properties of fluorescent proteins and dyes. SAMI-qSMLM afforded robust quantification. To efficiently detect endogenous proteins, we used fluorescent ligands that bind to a specific site on engineered antibody fragments. Both the density and nano-organization of membrane-bound epidermal growth factor receptors (EGFR, HER2, and HER3) were determined by a combination of SAMI, antibody engineering, and pair-correlation analysis. In breast cancer cell lines, we detected distinct differences in receptor density and nano-organization upon treatment with therapeutic agents. This new platform can improve molecular quantification and can be developed to study the local protein environment of intact cells.

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“…В опухолевых клетках различные представители семейства GPCRs могут взаимодействовать на этапе внутриклеточной передачи сигнала. Например, в клетках протокового рака поджелудочной железы имеет место образование гетеромеров OPRM1 и SSTR2 (somatostatin receptor 2), которое связано с метастатическим потенциалом опухоли и не наблюдается в нормальной панкреатической ткани [14]. SNP 118A>G гена OPRM1 сопровождается заменой аспарагина на аспартат в положении 40 аминокислотной структуры рецептора (Asn40Asp), что приводит к снижению экспрессии гена, повышению аффинитета к β-эндорфину и уменьшению чувствительности к экзогенным опиоидам [8].…”

Section: результаты и обсуждениеunclassified

Polymorphism of μ1-opioid receptor (OPRM1) gene may be of value in genesis of renal malignant neoplasm

et al. 2019

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Objective: to evaluate alleles distribution of single nucleotide polymorphism 118A>G of μ1-opioid receptor (OPRM1) gene in patients with renal neoplasm and benign diseases.Materials and methods. 100 consecutive patients after renal surgeries retrospectively divided into groups with neoplasm (n = 29) and benign diseases (n = 71).Results. The incidence of renal neoplasm was much higher in homozygous 118A patients than in AG + GG group (36.4 % vs. 14.7 %; p = 0.035).Conclusion. Single nucleotide polymorphism 118A>G OPRM1 gene may be of value in genesis of renal neoplasm.

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Molecular signatures of mu opioid receptor and somatostatin receptor 2 in pancreatic cancer

Cited by 31 publications

References 97 publications

Single molecule localization microscopy coupled with touch preparation for the quantification of trastuzumab-bound HER2

Single molecule localization microscopy coupled with touch preparation for the quantification of trastuzumab-bound HER2

A Platform To Enhance Quantitative Single Molecule Localization Microscopy

Polymorphism of μ1-opioid receptor (OPRM1) gene may be of value in genesis of renal malignant neoplasm

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