4 nya forskningsrapporter (1 överraskande)

 1. Stamceller för bröstcancerbehandling

Arton forskare från  Italienska och Australienska universitet har fått sina studier publicerade i Nature´s edition BMC,

Mitochondrial transfer from Adipose stem cells to breast cancer cells drives multi-drug resistance

Abstract

Background

Breast cancer (BC) is a complex disease, showing heterogeneity in the genetic background, molecular subtype, and treatment algorithm. Historically, treatment strategies have been directed towards cancer cells, but these are not the unique components of the tumor bulk, where a key role is played by the tumor microenvironment (TME), whose better understanding could be crucial to obtain better outcomes.

Methods

We evaluated mitochondrial transfer (MT) by co-culturing Adipose stem cells with different Breast cancer cells (BCCs), through MitoTracker assay, Mitoception, confocal and immunofluorescence analyses. MT inhibitors were used to confirm the MT by Tunneling Nano Tubes (TNTs). MT effect on multi-drug resistance (MDR) was assessed using Doxorubicin assay and ABC transporter evaluation. In addition, ATP production was measured by Oxygen Consumption rates (OCR) and Immunoblot analysis.

Materials and methods (urval)

Digital holographic microscopy

For kinetic dose–response assay, a Holomonitor M4 microscope (Phase Holographic Imaging AB, Lund, Sweden) was used. Cells were seeded at a density of 104 cells/well and then subjected to MCP before treatment with DOX (1 µM for MDA-MB.231; 2 µM for MCF-7). Cells were continuously monitored for 24 h, in time-lapse mode (every hour), at multiple positions in each well using a high-precision motorized stage. The Holomonitor App suite cell imaging software was used for image analysis, for the evaluation of the cell viability parameters (cell density = #cells/cm2; percentage of confluence = %confluence). The results are showed as changes relative to zero time-point, from each area and presented as mean ± SE for all the monitored areas in each well. The assay has been performed three times in triplicate.

Five view fields were analyzed for each sample. Statistical significance for Holomonitor data was determined by the Ordinary Two-Way Anova model, adjusted with the Sidak multiple comparison test. Unless otherwise specified statistical analysis were performed with GraphPad Prism9. Two-tailed p values < 0.05 were considered significant.

Subsequently, we focused our attention on the possible effect of ADM on ABC transporters activity. Preliminarily, we evaluated DOX cytotoxicity on BCCs after MCP. Indeed, it induced an increase of cell viability parameters monitored with Holomonitor (***P ≤ 0.001), also in hypoxia (**P ≤ 0.01). 

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2. Trombocyter i benmärgen

I en specialutgåva har 5 engelska forskare fått sina studier publicerade om möjligheten att påverka trombocytproduktionen (blodplättar) i benmärgen. Behovet av ökad produktion av dessa plättar infinner sig nästan alltid vid en cancerbehandling. Även jag fick substanser för att få igång min produktion när jag genomgick min cellgiftsbehandling.Det då cellgifterna skördade allt (nästan) i sin väg för att ha ihjäl mina cancerceller.

Pim Kinase Inhibition Disrupts CXCR4 Signalling in Megakaryocytes and Platelets by Reducing Receptor Availability at the Surface

Abstract

A key step in platelet production is the migration of megakaryocytes to the vascular sinusoids within the bone marrow. This homing is mediated by the chemokine CXCL12 and its receptor CXCR4. CXCR4 is also a positive regulator of platelet activation and thrombosis. Pim-1 kinase has been shown to regulate CXCR4 signalling in other cell types, and we have previously described how Pim kinase inhibitors attenuate platelet aggregation to CXCL12. However, the mechanism by which Pim-1 regulates CXCR4 signalling in platelets and megakaryocytes has yet to be elucidated. Using human platelets, murine bone marrow-derived megakaryocytes, and the megakaryocyte cell line MEG-01, we demonstrate that pharmacological Pim kinase inhibition leads to reduced megakaryocyte and platelet function responses to CXCL12, including reduced megakaryocyte migration and platelet granule secretion. Attenuation of CXCL12 signalling was found to be attributed to the reduced surface expression of CXCR4. The decrease in CXCR4 surface levels was found to be mediated by rapid receptor internalisation, in the absence of agonist stimulation. We demonstrate that pharmacological Pim kinase inhibition disrupts megakaryocyte and platelet function by reducing constitutive CXCR4 surface expression, decreasing the number of receptors available for agonist stimulation and signalling. These findings have implications for the development and use of Pim kinase inhibitors for the treatment of conditions associated with elevated circulating levels of CXCL12/SDF1α and increased thrombotic risk.

Materials and Methods (urval)

4.8. Live Cell migration/Motility Assay

MEG01 cells were plated in 24-well plates and allowed to attach for three days before non-adherent cells were washed off. Cells were serum-starved overnight in serum-free media, then cells were treated with AZD1208 (10 µM) or the vehicle (0.1% DMSO) for 24 h in the presence of 100 ng/mL CXCL12. Images were acquired at 37 °C and 5% CO2 using a Holomonitor M4 live cell imaging system (Phase Holographic Imaging, PHI, Lund, Sweden). Data analysis was performed using the HoloMonitor App Suite 3.5.1 (PHI AB, Lund, Sweden) to calculate average cell motility and migration. 

Figure 2. Pim kinase inhibition attenuates megakaryocyte migration to CXCL12. (A) MEG01 cells or (B) murine bone marrow-derived mature megakaryocytes were incubated with AZD1208 (10 µM) or the vehicle (0.1% DMSO) control for 30 min, prior to the addition of 100 ng/mL CXCL12. (A i) MEG01 cell migration and (A ii) motility in the presence of CXCL12 was recorded and measured over time (24 h) using a Holomonitor Live Cell Imaging system. (B) Primary megakaryocytes were added to transwell membrane inserts, and migration towards CXCL12 (300 ng/mL) was measured for 5 h using DAPI, counting the number of cells that passed through the transwell insert using a Celena Logos and 4x objective lens. (B i) Representative images and (B ii) quantified data are shown. (A,B) All quantified data points are shown (◦ white circles = vehicle, grey circles = + AZD1208)), n = 4, error bars are mean ± SEM, * p ≤ 0.05, ** p ≤ 0.01.

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3. Studie om uppkomst av KOL ( Kroniskt Obstruktiv Lungsjukdom)

I en specialutgåva har 10 svenska Lundaforskare publicerat sina studier om bindvävsceller (fibroblaster) i en KOL-lunga gentemot en frisk lunga.Tesen är att återskapa nya fibroblastceller i en KOL-lunga och få den fungerande igen.

Expression of Stress-Induced Genes in Bronchoalveolar Lavage Cells and Lung Fibroblasts from Healthy and COPD Subjects

Abstract

Chronic obstructive pulmonary disease (COPD) is commonly caused from smoking cigarettes that induce biological stress responses. Previously we found disorganized endoplasmic reticulum (ER) in fibroblasts from COPD with different responses to chemical stressors compared to healthy subjects. Here, we aimed to investigate differences in stress-related gene expressions within lung cells from COPD and healthy subjects. Bronchoalveolar lavage (BAL) cells were collected from seven COPD and 35 healthy subjects. Lung fibroblasts were derived from 19 COPD and 24 healthy subjects and exposed to cigarette smoke extract (CSE). Gene and protein expression and cell proliferation were investigated. Compared to healthy subjects, we found lower gene expression of CHOP in lung fibroblasts from COPD subjects. Exposure to CSE caused inhibition of lung fibroblast proliferation in both groups, though the changes in ER stress-related gene expressions (ATF6, IRE1, PERK, ATF4, CHOP, BCL2L1) and genes relating to proteasomal subunits mostly occurred in healthy lung fibroblasts. No differences were found in BAL cells. In this study, we have found that lung fibroblasts from COPD subjects have an atypical ER stress gene response to CSE, particularly in genes related to apoptosis. This difference in response to CSE may be a contributing factor to COPD progression.

2.3. The Effect of CSE on Cell Proliferation

To investigate the potential effect of CSE on lung fibroblasts, the proliferation of lung fibroblasts from healthy and COPD subjects was quantified using the HoloMonitor. We found that the proliferation of lung fibroblasts from both healthy and COPD subjects was inhibited during exposure with 5% CSE at 48 h (p = 0.05 in both, Figure 5), with no significant differences between COPD and healthy subjects.

Figure 5. Lung fibroblast proliferation during 48 h measured by HoloMonitor. The graph presents cell growth as % of time point 0. Lung fibroblasts from healthy subjects (n = 3) and COPD patients (n = 3) were investigated without (0% CSE) or with Cigarette smoke extract (5% CSE). † and ‡ = significantly different trend (p = 0.05) from 0 h to 48 h between 0 and 5% CSE within healthy and COPD subjects, respectively. Friedman’s test was used for statistical analysis of differences in trends over time between 0 and 5% CSE.

4.9. Analyses of Cell Proliferation Using HoloMonitor

Human lung fibroblasts were grown in 6-well plates (Sarstedt TC, Ref. 83,3920, Nümbrecht, Germany) for 24 h prior to CSE exposure and were seeded at a concentration of 10,000 cells/cm2. These cells were exposed to either 0% or 5% CSE and studied using the HoloMonitor M4 live cell imaging system (Phase Holographic Imaging, Lund, Sweden). For each sample, 4–7 focus points were set per well and exposure, and images were captured over a time period of 48-h. The average percentage of cell growth relative to the starting number of cells at the 0 h time point was calculated per individual.

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4. SARS (Covid-19?) relaterad forskning

Slutligen en överraskande studie från 8 indiska forskare. Överraskande i så måtto att HoloMonitor inte använts i SARS forskningen tidigare samt att Indien ser ut att ha köpt sitt första HoloMonitorsystem.

Forskarna ifråga är nästan barnsligt förtjusta i vad HoloMonitor-tekniken erbjuder,läs detta: 

Holography microscopy-based insights into SARS-CoV E protein-derived pentapeptide interactions with intestinal cells: 

 In this study, we investigated the impact of SARS-CoV E protein-derived pentapeptides on human intestinal cells using holography microscopy over the established MTT assay. 

Holomonitor microscopy, an innovative imaging technique that allows us to continuously monitor the live cellular responses to these nanostructured materials in real time, within their natural environment. The major advantage of this microscopy is its label-free and non-invasive nature; cells do not have to be stained or fixed, which might influence cell physiology. 

While the MTT assay, is invasive and provides an endpoint measurement of cell viability based on metabolic activity, it is a widely accepted method for detecting if cells are alive or dead and is limited in its ability to deliver detailed information about cellular health and mechanisms. 

The holography microscopy generates detailed data on cell volume, morphology, and confluence. 

These parameters provide deeper insights into cellular mechanisms and how they are affected by treatments over time. The approach we use is particularly helpful in our study area. 

It sheds light on the interaction between nanostructures and cells and what happens at the interface between them. 

We believe that understanding this interface is of utmost importance for actual therapeutic applications, which might rely in the future on the generation of hybrid nanostructures that can interface not just with cells but also with various tissues and organs of the human body.

Studien publicerades för 2 dagar sen och är betitlad: 

Multiscale Materials Engineering via Self-Assembly of Pentapeptide Derivatives from SARS CoV E Protein

Dessvärre inlåst,men under Supporting Information hittar man forskarnas text om HoloMonitor samt denna illustration vid användandet av det de kallar The state-of-the-art,dvs HoloMonitor.

man

Figure S37. Impact of peptides P1, P2, and P3 on the health and growth characteristics of intestinal epithelial SW620 cells over time. The state-of-the-art label-free live cell holography imaging was utilized to quantify cell count, confluence, and volume every 15 minutes over 72 hours following peptide treatment. (A) Time-lapse depiction of the effect of peptides on SW620 cell count illustrates the average normalized cell count over the specified intervals. (B) Time-lapse visualization of the effect of peptides on SW620 cell confluence. (C) Time-lapse illustration of peptides' effect on SW620 cell volume over the specified intervals.   

Tillägg 19/7

Mr google är behjälplig och verifierar indiernas användande av HoloMonitor i denna studie.

Min kommentar
Dessa 4 studier är alla med olika forskningsinnehåll med den gemensamma nämnaren HoloMonitor.

QPI-tekniken visar här tydligt hur användbar den är inom den bredare forskningen. Att QPI kommer bli en Golden Standard är förmodligen bara en tidsfråga. Begrunda studierna och se möjlig potential för X antal olika forskningsinriktningar med tekniken.

                                             Mvh the99

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