söndag 29 juni 2025

PHI - En viktig forskningsrapport

Trippelnegativ bröstcancer bör vara alla kvinnors fasa när det kommer till att diagnostisera bröstcancer.
Den trippla cancern är verkligen en ruskig best då den är extremt aggressiv, med jobbiga konsekvenser om den kan och får härja fritt. Upptäcks den för sent eller misstas för en vanlig bröstcancer är risken mycket stor att den metastaserar,dvs sprider sig. Vanligaste områdena då är till hjärnan,skelettet,lungorna och levern. Vad som utmärker den trippla cancern mot den vanliga är att den saknar 3 receptorer som finns på cancercellernas yta. Dessa är Östrogen,Progesteron och HER2.
Avsaknaden gör att vanliga cancermediciner man använder inte biter på besten,därav vikten av ett snabbt konstaterande. Eftersom den trippla är sanslöst aggressiv vill den sprida sig,och det fort. Kommer man in för kontroll av misstänkt bröstcancer görs en nålbiopsi som tar vävnadsprov och analyseras för att få besked om det går att utesluta trippelcancern. Svar på detta kan ta allt från 1 vecka till flera veckor.
Konstateras trippelcancer vet läkarna bättre vilka behandlingsmetoder som ska användas. Eftersom denna best är så aggressiv är tidsaspekten viktig. Har kvinnan gått med denna knöl i bröstet alltför länge innan hon går till läkare för undersökning är risken stor att den redan hunnit sprida sig. Då ser det tyvärr inte så ljust ut för vederbörande. Den andra tidsaspekten är att det gäller att få fram svar snabbt efter biopsin,ju snabbare man vet vad det handlar om ju större chans att rätt behandling sätts in. 
Mellan 10-15% diagnostiseras årligen med den trippla varianten av bröstcancer.

Det här var en utvikning om bröstcancer som då leder mig till dagens ämne: 
Nytt sätt att särskilja trippelnegativ bröstcancer gentemot den vanliga.

Amerikanska forskare vid University of Dearborn Michigan har studerat skillnaden mellan en "vanlig" bröstcancercellinje (MCF10A) och den trippla cancercellinjen (MDA-MB-231) morfologiskt (b.l.a storlek och form) och bägge celllinjernas resp utveckling. Under 24 timmar studerades dessa bägge cellinjer i en HoloMonitor. 
Med PHI´s mjukvara Holostudio samt AI fick man fram konkreta skillnader och förklaringar till resultaten. Forskningsrapporten publicerades i fredags 27/6 i det vetenskapliga organet Cancer Reports
Från studien betitlad 

Dissecting Morphological and Functional Dynamics of Non-Tumorigenic and Triple-Negative Breast Cancer Cell Lines Using PCA and t-SNE Analysis

kommer här några utdrag:

ABSTRACT

Background

Triple-negative breast cancer (TNBC) poses significant challenges due to its aggressive nature and lack of targeted therapies. Understanding the cellular behaviors of TNBC is crucial for developing effective treatments.

Aims

This study aims to compare the morphological characteristics of non-tumorigenic MCF10A and aggressive MDA-MB-231 TNBC cell lines using advanced analytical techniques.

Methods and Results

Advanced techniques such as Principal Component Analysis (PCA), t-Distributed Stochastic Neighbor Embedding (t-SNE), and digital holographic microscopy were utilized. Cellular features such as area, migration, motility, irregularity, and optical thickness were thoroughly analyzed over time. Our results revealed significant morphological differences between the MCF10A and MDA-MB-231 cell lines. Specifically, MDA-MB-231 cells displayed enhanced motility and a smaller, more variable size, attributes that may facilitate their invasive potential. In contrast, MCF10A cells exhibited larger sizes and more regular migration patterns, suggesting stability in structured tissue environments. Additionally, temporal analysis highlighted consistent phenotypic behaviors over time, with MDA-MB-231 cells demonstrating higher optical thickness and irregularity, indicating potential structural complexities associated with malignant transformation. Correlative analysis further confirmed these results by revealing connections between cell size, motility, and optical properties crucial for understanding cell behavior within their microenvironment.

Conclusion

The profound differences in cellular dynamics between MCF10A and MDA-MB-231 cell lines underscore the unique adaptive mechanisms of TNBC cells. Our study provides valuable insights into the cellular foundations of TNBC aggressiveness, offering a foundation for future research aimed at understanding the mechanistic underpinnings of TNBC progression and therapeutic targeting.

1 Introduction

Breast cancer remains one of the most common malignancies among women worldwide, presenting significant variability in its biological behavior and clinical outcomes. Understanding the cellular mechanisms underlying breast cancer phenotypes, particularly the aggressive triple-negative breast cancer (TNBC), is crucial for developing targeted therapies and improving prognosis  Morphological and functional analyses are critical for understanding cancer progression, as traits such as cell motility, migration patterns, and size directly influence the invasive and metastatic potential of cancer cells. By investigating these characteristics, researchers can identify cellular behaviors that differentiate aggressive cancer subtypes like TNBC from normal tissue, providing insights into their ability to adapt to and invade diverse microenvironments. 

This study focuses on comparing the morphological and functional characteristics of non-tumorigenic and tumorigenic breast cell lines, specifically the well-characterized MCF10A and MDA-MB-231 lines, to elucidate the cellular behaviors that drive malignancy and metastatic potential in TNBC. Despite significant advances in breast cancer research, the specific morphological and functional mechanisms that distinguish TNBC cells from non-tumorigenic counterparts, to the best of our knowledge, remain poorly understood. Addressing this gap is critical for identifying biomarkers and therapeutic targets that could mitigate TNBC's aggressive behavior. This study aims to fill this gap by leveraging advanced analytical techniques to uncover these distinctions in detail.

Employing techniques such as Principal Component Analysis (PCA) and t-Distributed Stochastic Neighbor Embedding (t-SNE), this study comprehensively analyzes the differences between these cell lines. PCA is a linear dimensionality reduction technique that simplifies complex datasets by identifying the main components contributing to variability. In contrast, t-SNE is a non-linear method that captures local and global structures within data, enabling the visualization of clustering patterns. These approaches provide an unbiased, high-dimensional perspective on cellular behavior, overcoming the limitations of traditional analyses that evaluate only one or two features at a time (e.g., plotting cell area versus migration distance in a 2D scatter plot). 
Utilization of Digital Holographic Microscopy (DHM), a label-free imaging technique, further enhances this approach by allowing real-time, non-invasive quantification of cellular morphology and dynamics, helping in preserving native cell behavior while at the same time generating richer datasets. Altogether, these advanced computational and imaging methods enable a deeper, more nuanced understanding of TNBC aggressiveness, revealing phenotypic heterogeneity that might be overlooked with conventional techniques.

2 Materials and Methods

2.1 Cell Culture and Digital Holographic Microscopy Analysis

Digital Holographic Microscopy (DHM) is a label-free, quantitative imaging technique that captures real-time cellular morphology and dynamics by recording interference patterns of coherent light passing through or reflected from cells. This approach enables the non-invasive study of live cells, preserving their natural behavior while providing high-resolution quantitative phase images. Unlike fluorescence microscopy, which often requires labeling that can perturb cellular behavior, DHM allows for longitudinal observations under near-physiological conditions. These characteristics make DHM particularly well-suited for studies requiring the tracking of cellular motility, migration, and structural changes over time. DHM operates by capturing interference patterns generated as coherent light passes through cells, which are then computationally reconstructed to provide both phase and amplitude information. The resulting quantitative phase images enable measurement of cellular morphology and dynamic behaviors with high spatial and temporal resolution. Holominitor uses Fourier-based digital holography to extract key features, following established methodologies for refractometry and quantitative analysis of adherent cells.

In this study, DHM was employed to investigate the morphological and functional differences between non-tumorigenic MCF10A and aggressive triple-negative breast cancer MDA-MB-231 cell lines. The ability of DHM to provide label-free, real-time imaging allowed for a comprehensive analysis of key parameters such as cell size, motility, irregularity, and optical thickness without disrupting cellular integrity. These unique features of DHM facilitated the temporal tracking of cellular behaviors and provided robust datasets for subsequent high-dimensional analysis using PCA and t-SNE. MCF10A (ATCC CRL-10317) and MDA-MB-231 (ATCC HTB-26) were seeded at approximately 50% confluency in MEBM Basal Medium supplemented with BPE, hEGF, Insulin, hydrocortisone, GA-1000 (Lonza, Basel, Switzerland) and RPMI1640, 10% FBS, 5% Penicillin/Streptomycin, Insulin (1 mg/mL, Gibco, New York, USA) at 37°C with 5% CO2. The study employed a HoloMonitor M4, manufactured by Phase Holographic Imaging (PHI, Lund, Sweden)
The Hstudio software package from PHI enabled time-lapse phase imaging, image manipulation, segmentation, and data examination. Images were captured at 15-min intervals, starting 24 h after cell seeding. Temporal changes in cellular morphology, including features such as area, motility, and irregularity, were analyzed using time-lapse data generated by Digital Holographic Microscopy (DHM). Quantitative measurements were extracted using HStudio software and subsequently processed in Python for statistical analysis. Temporal patterns were assessed through feature-specific scatter plots, and independent two-sample t-tests were applied to evaluate significant differences over time between the cell lines. PCA and t-SNE were used to visualize temporal clustering and identify trends in high-dimensional data. The images from each well were divided into distinct sections, identified, and the number of cells was determined using the Cell Count function of the HStudio software. For the tracking as mentioned above assays, cells were cultured following standard conditions and seeded in 24-well plates (Sarstedt, Hildesheim, Germany).

2.2 Data Collection and Processing

The current study thoroughly compared MCF10A and MDA-MB-231 cell lines, which are representative of healthy mammary epithelial cells and aggressive triple-negative breast cancer cells, respectively. Our approach involved several steps. The HoloMonitor M4 software collected multiple cellular features, including area, irregularity, migration, motility, migration directness, and optical thickness. These specific features were selected for analysis based on their biological relevance to cell morphology and motility, their statistical variability between the two cell lines, and to avoid redundancy. 

HoloMonitor visar här A. som är vanliga bröstcancerceller och B. den trippla varianten.

Min kommentar
Jag ska försöka avhålla mig från min vanliga kanske överentusiastiska tirad om hur jäv..a bra HoloMonitor är vid cancerforskning (svårt) och istället konstatera att på 24 timmar + ev tid för analysarbetet har PHI`s HoloMonitor gett läkarkåren ett snabbare sätt att diagnostisera en trippelnegativ bröstcancer mot den traditionella metoden med biopsi + labbande som tar från 1 vecka till flera veckor (månad?). Utvecklar PHI detta och får till alla godkännanden för kliniskt användande av HoloMonitor och såklart även CellSync hägrar en bamsebamse-marknad upphöjt till 10. Tänk alla sjukhus och kliniker världen över om de fick tillgång till ett instrument som kortar ner svarstiden från veckor till dagar. Tänk hur många fler kvinnor som då får en bättre chans att överleva denna jäv...a best. 
(Ok,jag klarade kanske inte helt att avhålla mig från den överentusiastiska stilen. 😎)
                                          Mvh the99

Som service till alla HoloMonitornyfikna forskare : HoloMonitor Demo

2 kommentarer:

  1. Bra där 👍🏻 the99🥰

    SvaraRadera
  2. Hamnade hos NIH också https://pmc.ncbi.nlm.nih.gov/articles/PMC12203836/

    SvaraRadera