App-based support for breast cancer patients to reduce psychological distress during therapy and survivorship – a multicentric randomized controlled trial

IntroductionThe negative impact of unmanaged psychological distress on quality of life and outcome in breast cancer survivors has been demonstrated. Fortunately, studies indicate that distress can effectively be addressed and even prevented using evidence-based interventions. In Germany prescription-based mobile health apps, known as DiGAs (digital health applications), that are fully reimbursed by health insurances, were introduced in 2020. In this study, the effectiveness of an approved breast cancer DiGA was investigated: The personalized coaching app PINK! Coach supports and accompanies breast cancer patients during therapy and follow-up.MethodsPINK! Coach was specifically designed for breast cancer (BC) patients from the day of diagnosis to the time of Follow-up (aftercare). The app offers individualized, evidence-based therapy and side-effect management, mindfulness-based stress reduction, nutritional and psychological education, physical activity tracking, and motivational exercises to implement lifestyle changes sustainably in daily routine. A prospective, intraindividual RCT (DRKS00028699) was performed with n = 434 patients recruited in 7 German breast cancer centers from September 2022 until January 2023. Patients with BC were included independent of their stage of diseases, type of therapy and molecular characteristics of the tumor. Patients were randomized into one of two groups: The intervention group got access to PINK! over 12 weeks; the control group served as a waiting-list comparison to “standard of care.” The primary endpoint was psychological distress objectified by means of Patient Health Questionnaire-9 (PHQ-9). Subgroups were defined to investigate the app’s effect on several patient groups such as MBC vs. EBC patients, patients on therapy vs. in aftercare, patients who received a chemotherapy vs. patients who did not.ResultsEfficacy analysis of the primary endpoint revealed a significant reduction in psychological distress (least squares estimate -1.62, 95% confidence interval [1.03; 2.21]; p<0.001) among intervention group patients from baseline to T3 vs, control group. Subgroup analysis also suggested improvements across all clinical situations.ConclusionPatients with breast cancer suffer from psychological problems including anxiety and depression during and after therapy. Personalized, supportive care with the app PINK! Coach turned out as a promising opportunity to significantly improve psychological distress in a convenient, accessible, and low-threshold manner for breast cancer patients independent of their stage of disease (EBC/MBC), therapy phase (aftercare or therapy) or therapy itself (chemotherapy/other therapy options). The app is routinely available in Germany as a DiGA. Clinical Trial Registration: DRKS Trial Registry (DRKS00028699)

Promoter methylation-associated loss of ID4 expression is a marker of tumour recurrence in human breast cancer

<p>Abstract</p> <p>Background</p> <p>Inhibitor of DNA binding/Inhibitor of differentiation 4 (<it>ID4</it>) is a critical factor for cell proliferation and differentiation in normal vertebrate development. <it>ID4</it> has regulative functions for differentiation and growth of the developing brain. The role of <it>ID1</it>, <it>ID2</it> and <it>ID3</it> are expected to be oncogenic due to their overexpression in pancreatic cancer and colorectal adenocarcinomas, respectively. Aside from these findings, loss of <it>ID3</it> expression was demonstrated in ovarian cancer. The aim of the present study was to reveal the factual role of <it>ID4</it> in carcinogenesis in more detail, since its role for the pathogenesis of human breast cancer has been discussed controversially, assigning both oncogenic and tumour suppressive functions. </p> <p>Methods</p> <p><it>ID4</it> promoter methylation, <it>ID4</it> mRNA expression and <it>ID4</it> protein expression were analysed in primary human breast cancer specimens using methylation-specific PCR (MSP) (n=170), semiquantitative realtime RT-PCR (n=46) and immunhistochemistry (n=3), respectively. In order to demonstrate a functional association of <it>ID4</it> promoter methylation with its gene silencing, we performed DNA demethylation analysis with four human breast cell lines using MSP and semiquantitative realtime RT-PCR. In addition, we performed correlations of <it>ID4</it> promoter methylation with <it>ID4</it> mRNA and <it>ID4</it> protein expression in matched samples of breast tumour and corresponding normal tissue. We carried out statistical analyses in order to find correlations between <it>ID4</it> promoter methylation and clinicopathological parameters. </p> <p>Results</p> <p>Frequent <it>ID4</it> promoter methylation was observed in primary breast cancer samples (69%, 117/170). We found a tight correlation (P<0.0001) between <it>ID4</it> promoter methylation and loss of <it>ID4</it> expression in primary breast cancer 3 specimens. Demethylating treatment with breast cancer cell lines was associated with clear ID4 mRNA re-expression. Tumours with <it>ID4</it> promoter methylation showed distinct loss of <it>ID4</it> expression on both transcription and protein level. Interestingly, <it>ID4</it> promoter methylation was a factor for unfavourable recurrence-free survival (P=0.036) and increased risk for lymph node metastasis (P=0.030). </p> <p>Conclusion</p> <p>ID4 is indeed a novel tumour suppressor gene in normal human breast tissue and is epigenetically silenced during cancer development, indicating increased risk for tumour relapse. Thus, <it>ID4</it> methylation status could serve as a prognostic biomarker in human breast cancer.</p

Elemental localization of the von Economo neuron and fork neuron in the insular cortex in humans and macaque monkeys

The cerebral cortex of mammalian species is parcellated into discrete areas, or modules. Some of these modules share common features amongst most mammals; however, between higher and lower mammalian species, deviations in size, subdivisions, cellular features and connectivity of these modules exist. The cortex of higher mammals like primates, has expanded and has subdivided many times resulting in a complex neural system built of smaller modules acting in concert and forming functional networks. In line with this, in primates, the insular cortex has been shown to consist of smaller, unique modules, each having a specific structure and function. Furthermore, the research presented here, identified that in the anterior agranular and the dysgranular insula of macaque monkeys one of each of these architectonic area harbors two specialized neuronal morphotypes, the von Economo neurons (VEN) and the fork neurons (FN), whilst in the human anterior insula there are five distinct architectonic areas hosting these neurons. More than a century ago, Brodmann defined this kind of containment of specialized neuronal morphotypes with a specific cytoarchitectonic area of the primate cerebral cortex as “elemental localization”, because it suggests shared evolutional, developmental, and functional features between the specialized neurons and their host area. The discovery of novel elemental localizations within the primate brain 1) supports the theory that the ventral anterior insular cortex as well as the adjacent orbital prefrontal cortex are heterogeneously organized into smaller areas rather than being lumped together into a larger, homogeneous periallocortical (agranular) sector, and 2) provides an exclusive experimental advantage for the examination of the VEN areas and its two specialized neuronal morphotypes, VEN and FN, which are linked to internal bodily state representation and neuropsychiatric disorders

RNA sequencing of glioblastoma tissue slice cultures reveals the effects of treatment at the transcriptional level

One of the major challenges in cancer research is finding models that closely resemble tumors within patients. Human tissue slice cultures are a promising approach to provide a model of the patient's tumor biology ex vivo. Recently, it was shown that these slices can be successfully analyzed by whole transcriptome sequencing as well as automated histochemistry, increasing their usability as preclinical model. Glioblastoma multiforme (GBM) is a highly malignant brain tumor with poor prognosis and little is known about its genetic background and heterogeneity regarding therapy success. In this study, tissue from the tumors of 25 patients with primary GBM was processed into slice cultures and treated with standard therapy (irradiation and temozolomide). Total RNA sequencing and automated histochemistry were performed to enable analysis of treatment effects at a transcriptional and histological level. Slice cultures from long‐term survivors (overall survival [OS] > 24 months) exhibited more apoptosis than cultures from patients with shorter OS. Proliferation within these slices was slightly increased in contrast to other groups, but not significantly. Among all samples, 58 protein‐coding genes were upregulated and 32 downregulated in treated vs. untreated slice cultures. In general, an upregulation of DNA damage‐related and cell cycle checkpoint genes as well as enrichment of genotoxicity pathways and p53‐dependent signaling was found after treatment. Overall, the current study reproduces knowledge from former studies regarding the feasibility of transcriptomic analyses and automated histology in tissue slice cultures. We further demonstrate that the experimental data merge with the clinical follow‐up of the patients, which improves the applicability of our model system

Translocator protein (18 kDa) (TSPO) is expressed in reactive retinal microglia and modulates microglial inflammation and phagocytosis

Background: The translocator protein (18 kDa) (TSPO) is a mitochondrial protein expressed on reactive glial cells and a biomarker for gliosis in the brain. TSPO ligands have been shown to reduce neuroinflammation in several mouse models of neurodegeneration. Here, we analyzed TSPO expression in mouse and human retinal microglia and studied the effects of the TSPO ligand XBD173 on microglial functions. Methods: TSPO protein analyses were performed in retinoschisin-deficient mouse retinas and human retinas. Lipopolysaccharide (LPS)-challenged BV-2 microglial cells were treated with XBD173 and TSPO shRNAs in vitro and pro-inflammatory markers were determined by qRT-PCR. The migration potential of microglia was determined with wound healing assays and the proliferation was studied with Fluorescence Activated Cell Sorting (FACS) analysis. Microglial neurotoxicity was estimated by nitrite measurement and quantification of caspase 3/7 levels in 661 W photoreceptors cultured in the presence of microglia-conditioned medium. The effects of XBD173 on filopodia formation and phagocytosis were analyzed in BV-2 cells and human induced pluripotent stem (iPS) cell-derived microglia (iPSdM). The morphology of microglia was quantified in mouse retinal explants treated with XBD173. Results: TSPO was strongly up-regulated in microglial cells of the dystrophic mouse retina and also co-localized with microglia in human retinas. Constitutive TSPO expression was high in the early postnatal Day 3 mouse retina and declined to low levels in the adult tissue. TSPO mRNA and protein were also strongly induced in LPS-challenged BV-2 microglia while the TSPO ligand XBD173 efficiently suppressed transcription of the pro-inflammatory marker genes chemokine (C-C motif) ligand 2 (CCL2), interleukin 6 (IL6) and inducible nitric oxide (NO)-synthase (iNOS). Moreover, treatment with XBD173 significantly reduced the migratory capacity and proliferation of microglia, their level of NO secretion and their neurotoxic activity on 661 W photoreceptor cells. Furthermore, XBD173 treatment of murine and human microglial cells promoted the formation of filopodia and increased their phagocytic capacity to ingest latex beads or photoreceptor debris. Finally, treatment with XBD173 reversed the amoeboid alerted phenotype of microglial cells in explanted organotypic mouse retinal cultures after challenge with LPS. Conclusions: These findings suggest that TSPO is highly expressed in reactive retinal microglia and a promising target to control microglial reactivity during retinal degeneration

Promoter methylation-associated loss of expression is a marker of tumour recurrence in human breast cancer-0

Ion-specific PCR (MSP) primers used for this study were positioned within the central CpG island (-615 until +139) near the TSS, detecting an amplicon indicated by the black box. The PCR product size of the unmethylated promoter sequence is 161 bp and similar in size to the product achieved with primers indicating methylated promoters (157 bp). The primers for the U reaction cover the bases -194 until -166 and -60 until -33. The primers for the M reaction cover the bases -192 until -166 and -60 until -35. All positions are relative to the TSS. . Cells were incubated with 5-aza-2'-deoxycytidine (DAC) and trichostatin A (TSA) for 72 h and 24 h, respectively. For each cell line, the methylation status and the fold change (FC) of mRNA re-expression are shown. All methylated cell lines (BT20, MCF7 and T47D) restored expression after demethylating treatment. MDA-MB231 remains unmethylated in the promoter and exhibits only a marginal increase of mRNA expression after DAC/TSA treatment. . MSP results from nine representative patients (#) are shown. DNA bands in lanes labelled with U indicate PCR products amplified with primers recognising the unmethylated promoter sequence. DNA bands in lanes labelled with M represent products amplified with primers specific for methylated alleles. Human breast cancer cell lines BT20 and MDA-MB321 were used as positive controls for methylated and unmethylated ID4 promoter sequences, as described previously [20]. Water was used as non template control (NTC).<p><b>Copyright information:</b></p><p>Taken from "Promoter methylation-associated loss of expression is a marker of tumour recurrence in human breast cancer"</p><p>http://www.biomedcentral.com/1471-2407/8/154</p><p>BMC Cancer 2008;8():154-154.</p><p>Published online 30 May 2008</p><p>PMCID:PMC2435120.</p><p></p

Promoter methylation-associated loss of expression is a marker of tumour recurrence in human breast cancer-3

Ion-specific PCR (MSP) primers used for this study were positioned within the central CpG island (-615 until +139) near the TSS, detecting an amplicon indicated by the black box. The PCR product size of the unmethylated promoter sequence is 161 bp and similar in size to the product achieved with primers indicating methylated promoters (157 bp). The primers for the U reaction cover the bases -194 until -166 and -60 until -33. The primers for the M reaction cover the bases -192 until -166 and -60 until -35. All positions are relative to the TSS. . Cells were incubated with 5-aza-2'-deoxycytidine (DAC) and trichostatin A (TSA) for 72 h and 24 h, respectively. For each cell line, the methylation status and the fold change (FC) of mRNA re-expression are shown. All methylated cell lines (BT20, MCF7 and T47D) restored expression after demethylating treatment. MDA-MB231 remains unmethylated in the promoter and exhibits only a marginal increase of mRNA expression after DAC/TSA treatment. . MSP results from nine representative patients (#) are shown. DNA bands in lanes labelled with U indicate PCR products amplified with primers recognising the unmethylated promoter sequence. DNA bands in lanes labelled with M represent products amplified with primers specific for methylated alleles. Human breast cancer cell lines BT20 and MDA-MB321 were used as positive controls for methylated and unmethylated ID4 promoter sequences, as described previously [20]. Water was used as non template control (NTC).<p><b>Copyright information:</b></p><p>Taken from "Promoter methylation-associated loss of expression is a marker of tumour recurrence in human breast cancer"</p><p>http://www.biomedcentral.com/1471-2407/8/154</p><p>BMC Cancer 2008;8():154-154.</p><p>Published online 30 May 2008</p><p>PMCID:PMC2435120.</p><p></p

Promoter methylation-associated loss of expression is a marker of tumour recurrence in human breast cancer-1

Breast cancer. The Y axis indicates the factor of mRNA downregulation in breast cancer specimens (n = 46) relative to a normal breast standard (a pool of 12 normal breast tissue samples) as the fold change (FC) N/T. Unmethylated tumours exhibited ID4 expression (median FC = 1.3) very similar to normal breast cells. In contrast, methylated breast cancer specimens displayed an increased loss of expression (median FC = 12.3). Horizontal lines: group medians; boxes: 25–75% quartiles; vertical lines: range, peak and minimum. . Distribution of time (months) and tumour-related death among 115 breast cancer patients with positive (lower graph) or negative (upper graph) promoter methylation state is shown. Patients harbouring an -methylated tumour have an estimated mean RFS time of 80 months (95% confidence interval: 67–93 months) compared with 101 months (95% confidence interval: 87–115 months) for patients without tumour methylation. See text for details.<p><b>Copyright information:</b></p><p>Taken from "Promoter methylation-associated loss of expression is a marker of tumour recurrence in human breast cancer"</p><p>http://www.biomedcentral.com/1471-2407/8/154</p><p>BMC Cancer 2008;8():154-154.</p><p>Published online 30 May 2008</p><p>PMCID:PMC2435120.</p><p></p

Promoter methylation-associated loss of expression is a marker of tumour recurrence in human breast cancer-2

Er and corresponding normal tissue (matched pairs a, b, c). Normal tissues are labelled with N. Their corresponding tumours are labelled with T. For each sample, promoter methylation, mRNA expression and ID4 immunohistochemical staining are shown (original magnification: 400×). The Fold change (FC) of mRNA downregulation is calculated by the expression ratio of normal (N) and tumourous tissue (T) for each matched pair (a, b, c). Matched pair a: Unmethylated tumour showed very similar mRNA and protein expression compared to the corresponding normal breast tissue. Matched pairs b and c: Methylated tumours exhibited explicit loss of mRNA and ID4 protein in comparison to the corresponding normal breast tissues (N). See text for details.<p><b>Copyright information:</b></p><p>Taken from "Promoter methylation-associated loss of expression is a marker of tumour recurrence in human breast cancer"</p><p>http://www.biomedcentral.com/1471-2407/8/154</p><p>BMC Cancer 2008;8():154-154.</p><p>Published online 30 May 2008</p><p>PMCID:PMC2435120.</p><p></p