Quantitative chromogenic immunohistochemical image analysis in cellprofiler software

Tollemar, Victor; Tudzarovski, Nikolce; Boberg, Erik; Andrén, Anton Törnqvist; Al-Adili, A.; Blanc, Katarina Le; Legert, Karin Garming; Bottai, Matteo; Warfvinge, Gunnar; Sugars, Rachael V.

doi:10.1002/cyto.a.23575

Cited by 14 publications

(14 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The advantage of using imaging analyses for scoring is that it is more objective and quantitative than manual assessment [34]. We found that patients with high ELTD1 staining in their primary tumor vasculature had a greater benefit from sunitinib treatment in terms of a prolonged PFS.…”

Section: Discussionmentioning

confidence: 84%

Tumor endothelial ELTD1 as a predictive marker for treatment of renal cancer patients with sunitinib

et al. 2020

View full text Add to dashboard Cite

Background: Patients with metastatic renal cell cancer (mRCC) are commonly treated with the tyrosine kinase inhibitor sunitinib, which blocks signalling from vascular endothelial growth factor (VEGF) -and platelet-derived growth factor-receptors, inhibiting development of new blood vessels. There are currently no predictive markers available to select patients who will gain from this treatment. Epidermal growth factor, latrophilin and seven transmembrane domain-containing protein 1 (ELTD1) is up-regulated in tumor endothelial cells in many types of cancer and may be a putative predictive biomarker due to its association with ongoing angiogenesis. Methods: ELTD1, CD34 and VEGF receptor 2 (VEGFR2) expressions were analysed in tumor vessels of renal cancer tissues from 139 patients with mRCC using immunohistochemistry. Ninety-nine patients were treated with sunitinib as the first or second-line therapy. Early toxicity, leading to the termination of the treatment, eliminated 22 patients from the analyses. The remaining (n = 77) patients were included in the current study. In an additional analysis, 53 sorafenib treated patients were evaluated. Results: Patients with high ELTD1 expression in the tumor vasculature experienced a significantly better progression free survival (PFS) with sunitinib treatment as compared to patients with low ELTD1 expression (8 versus 5.5 months, respectively). The expression level of CD34 and VEGFR2 showed no correlation to sunitinib response. In sorafenib treated patients, no association with ELTD1 expression and PFS/OS was found. Conclusions:Our results identify tumor vessel ELTD1 expression as a positive predictive marker for sunitinibtreatment in patients suffering from mRCC. The negative results in the sorafenib treated group supports ELTD1 being a pure predictive and not a prognostic marker for sunitinib therapy.

show abstract

Section: Discussionmentioning

confidence: 84%

Tumor endothelial ELTD1 as a predictive marker for treatment of renal cancer patients with sunitinib

et al. 2020

View full text Add to dashboard Cite

show abstract

“…CD163 immunolocalization was quantified using the open‐source CellProfiler software version 2 (http://www.cellprofiler.org). Total stained pixel area per image was calculated on the segmented identified objects and linear generalized estimating equations, to quantify CD163 expression within the VF tissue . Results are reported as total pixel area with 95% confidence intervals.…”

Section: Methodsmentioning

confidence: 99%

Mesenchymal stromal cells modulate tissue repair responses within the injured vocal fold

et al. 2019

View full text Add to dashboard Cite

Objectives/Hypothesis: This study aimed to determine whether local injection of human mesenchymal stromal cells (MSC) could modulate the early inflammatory response within injured vocal folds (VFs) to promote wound-healing processes.Study Design: Experimental xenograft model. Methods: VF injury was surgically induced by bilateral resection of the lamina propria of rabbits, and MSC were immediately injected into the injured area of both VFs. Animals were sacrificed on days 2, 4, and 24. Histological analyses were performed by hematoxylin and eosin, Masson's Trichrome, and elastin staining. Cell death was visualized by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and the M2 macrophage marker, CD163, detected by immunohistochemistry. Persistence of injected MSC was evaluated by fluorescent in situ hybridization (FISH). Quantitative polymerase chain reaction was performed on the contralateral VF.Results: Histological examination at days 2 and 4 indicated that MSC were able to reduce tissue inflammation, with gene expression analysis confirming a significant reduction of proinflammatory markers, interleukin (IL)-1β, and IL-8. FISH demonstrated low-level persistence of injected MSC at both time points, and TUNEL confirmed localized cell death at the injury site. Increased levels of CD163+ anti-inflammatory macrophages indicated a change in the immune milieu, supporting wound resolution. Evidence of a more organized collagen matrix suggests that MSC may enhance the production of a functional repair tissue after injury, despite their low-level persistence within the tissue.Conclusions: This study demonstrates that MSC are able to positively modulate the early wound-healing response through resolution of the inflammatory phase and promotion of tissue repair.

show abstract

“…The data was extracted from CellProfiler using customized Python code through Anaconda Software (Version 2-2.4.0). CellProfiler has been previously used and validated for both fluorescence and chromogenic localization by other research laboratories (McQuin et al, 2018;Tollemar et al, 2018). We further validated the accuracy of CellProfiler by comparing the counts from 26 ROIs between two experienced counters (manually) and CellProfiler.…”

Section: Automated Cell Countsmentioning

confidence: 98%

Calretinin and calbindin architecture of the midline thalamus associated with prefrontal–hippocampal circuitry

et al. 2020

View full text Add to dashboard Cite

The midline thalamus bi-directionally connects the medial prefrontal cortex (mPFC) and hippocampus (HC) creating a unique cortico-thalamo-cortico circuit fundamental to memory and executive function.While the anatomical connectivity of midline thalamus has been thoroughly investigated, little is known about its cellular organization within each nucleus. Here we used immunohistological techniques to examine cellular distributions in the midline thalamus based on the calcium binding proteins parvalbumin (PV), calretinin (CR), and calbindin (CB). We also examined these calcium binding proteins in a population of reuniens cells known to project to both mPFC and HC using a dual fluorescence retrograde adenoassociated virus (AAV) based tracing approach. These dual reuniens mPFC-HC projecting cells, in particular, are thought to be important for synchronizing mPFC and HC activity. First, we confirmed the absence of PV + neurons in the midline thalamus. Second, we found a common pattern of CR + and CB + cells throughout midline thalamus with CR + cells running along the nearby third ventricle (3V) and penetrating the midline. CB + cells were consistently more lateral and toward the middle of the dorsalventral extent of the midline thalamus. Notably, single-labeled CR + and CB + zones were partially overlapping and included dual-labeled CR + /CB + cells. Within RE, we also observed a CR and CB subzone specific diversity. Interestingly, dual mPFC-HC projecting neurons in RE expressed none of the calcium binding proteins examined, but were contained in nests of CR + and CB + cells. Overall, the midline thalamus was well organized into CR + and CB + rich zones distributed throughout the region, with dual mPFC-HC projecting cells in reuniens representing a unique cell population. These results provide a cytoarchitectural organization in the midline thalamus based on calcium binding protein expression, and sets the stage for future cell-type specific interrogations of the functional role of these different cell populations in mPFC-HC interactions..

show abstract

Quantitative chromogenic immunohistochemical image analysis in cellprofiler software

Cited by 14 publications

References 33 publications

Tumor endothelial ELTD1 as a predictive marker for treatment of renal cancer patients with sunitinib

Tumor endothelial ELTD1 as a predictive marker for treatment of renal cancer patients with sunitinib

Mesenchymal stromal cells modulate tissue repair responses within the injured vocal fold

Calretinin and calbindin architecture of the midline thalamus associated with prefrontal–hippocampal circuitry

Contact Info

Product

Resources

About