Ny forskningsrapport från Kina & Australien

I ett forskningsprojekt där forskare från 10 olika institut/universitet medverkar har info nu kommit om projektets innehåll och resultat.Amerikanska NIH med dess National Library of Medicine står bakom publiceringen.

Forskarna har gett sig på den svårbotade cancerformen trippelnegativ bröstcancer och studerat hur dess cancerceller svarar i en (förhållandevis) ny behandlingsform/terapi. Nämligen CAP = Cold Atmospheric Plasma.

CAP beskrivs enligt följande i en studie i Nature : CAP is an ionized near-room temperature gas, composed of reactive species, neutral particles and molecules, electrons and other physical factors such as electromagnetic field, weak ultraviolet radiation and weak heating effect. CAP has been widely used in many branches of modern medicine, including wound healing and sterilization. Over the past decade, CAP showed the promising application in the cancer treatment. CAP can effectively and selectively kill dozens of cancer cell lines in vitro through a direct CAP treatment on the cells or through an indirect CAP treatment on the medium or other biologically adaptable solutions which will be further used to affect the growth of cancer cells.

Vidare förklaring hur CAP fungerar rent praktiskt vågar jag mig inte på. Tekniken är tämligen invecklad,men det finns flera studier som visar på dess effektivitet. 

Men till forskningsprojektet och deras studie.

Cold atmospheric plasmas target breast cancer stemness via modulating AQP3-19Y mediated AQP3-5K and FOXO1 K48-ubiquitination

Abstract

Cold atmospheric plasma (CAP) is selective against many cancers with little side effect, yet its molecular mechanism remains unclear. Through whole transcriptome sequencing followed by assays in vitro, in vivo and using clinical samples, we propose CAP as a promising onco-therapy targeting cancer stemness via the AQP3/FOXO1 axis. CAP-generated reactive species penetrated cells via AQP3 and suppressed RPS6KA3, a shared kinase of AQP3 and FOXO1. Reduced AQP3-19Y phosphorylation suppressed SCAF11-mediated AQP3-5K K48-ubiquitination that led to sabotaged FOXO1 stability. Inhibited FOXO1 phosphorylation retarded its regulatory activities in maintaining cancer stemness including ALDH1 and IL6. Enhanced anti-cancer efficacy was observed through combining CAP with Atorvastatin in vitro and in vivo. We propose CAP as a 'selective' onco-therapeutic against cancer stemness, with the AQP3/FOXO1 axis being one molecular mechanism. We report SCAF11 as an E3 ubiquitin ligase of both AQP3 and FOXO1, identify AQP3-5K as an AQP3 K48-ubiquitination site, and emphasize the essential role of AQP3-19Y in this process. We reposition Atorvastatin into the onco-therapeutic portfolio by synergizing it with CAP towards enhanced efficacy. We anticipate the efficacy of CAP in targeting malignancies of high stemness alone or as an adjuvant therapy towards the hope of ultimate cancer cure.

Introduction

Cell phenotype switching between distinct states in response to environmental perturbations and mutational rewiring of the gene regulatory network is fundamental to cancer development and progression. Cancer therapy generally seeks to exploit this switching mechanism to force cancer cells into the apoptotic state. However, random and essentially uncontrollable transitions of highly-stressed surviving cells into the cancer stem cell (CSC) state often cause the failure of many therapeutic strategies.

Breast cancer is comprised of heterogeneous cell cohorts, with the triple negative breast cancer (TNBC) subtype being one of the most difficult to treat as they are easily attracted in the CSC state that defies effective therapeutic approach with little side effect. Close parallels have been made at the transcriptomic level between breast CSC and TNBC cell lines that fall into the claudin-low/basal B/mesenchymal molecular subgroup, enabling us to explore novel therapeutics and the molecular mechanisms against TNBCs using cell lines.

Cold atmospheric plasma (CAP), composed of reactive oxygen and nitrogen species (RONS) and electric fields, features multi-modal effects and can be generated via a range of device configurations, e.g., dielectric barrier discharge, plasma jet and plasma torch. CAP has been used as diverse medical therapies such as wound healing, sterilization, dental and dermatological treatments, as well as showcased its safety and efficacy in resolving cancers of, e.g., breast^, prostate, bladder, brain due to synergistic actions of its varied reactive components. Specifically, these reactive species interact with cancer cell surface to selectively arrest cancer cells at various death states such as immunogenic cell death, apoptosis cell cycle arrest and autophagy by relaying a series of signalings. Besides intensive preclinical efforts, the first clinical trial using CAP as an oncotherapy had been issued on 30 July 2019 and completed on 14 April 2021 in USA (NCT04267575) with success. Despite its immense translational potential as a first-line or adjuvant anti-cancer therapy, the mechanisms that enable safe, multi-modal efficacy of CAP against malignant cancers remain unclear.

Using TNBC cells as the model of malignant cells with high stemness, we aimed to explore the potential impact of CAP on cancer stemness, underlying molecular mechanism and possible drug synergies towards enhanced onco-therapeutic outcome in this study. Through whole transcriptome sequencing followed by assays in vitro, in vivo and using clinical samples, we propose CAP as a 'selective' onco-therapy targeting CSCs via the AQP3/FOXO1 axis. We report AQP3-5K K48-ubiquitination via SCAF11 with AQP3-19F phosphorylation being essential, and demonstrate synergies between CAP and Atorvastatin towards enhanced anti-cancer efficacy. We anticipate the feasibility of CAP in targeting other highly plastic cancers especially those lack safe cure, and its long-term success in synergizing with, e.g., immune- and chemo-therapies via arresting CSCs together with bulk tumor cells.

Under avsnittet Methods får vi bekräftelse på forskarnas användande av HoloMonitor.

HoloMonitor imaging

Cells were plated in 96-well plates at 5000 cells/well for 24 h before real-time monitoring. When the cell density was around 30%, the digital holograms of cells were set up using the HoloMonitor M4 Digital Holography Cytometer (Phase Holographic Imaging PHI AB, Lund, Sweden). The results were calculated using Hstudio M4 software (Phase Holographic Imaging PHI AB, Lund, Sweden).

 HoloMonitor imaging (Fig. ​Fig.11D) showed that CAP-activated medium (PAM) substantially reduced the random mobility of SUM159PT TNBC cells in terms of both migration distance (p=2.56E-5) and speed (p=1.9E-3)

CAP is selective against cancer stemness via the AQP3/FOXO1 axis. (A) Cell viabilities of different breast cancer cells, (B) HoloMonitor results on the distance and speed of SUM159PT cells, (C) glycolysis and (D) mitochondria ATP production rates, (E) ALDH⁺ cell cohort percentages, and (F) self-renew abilities of different breast cancer cells with and without CAP treatment. (G) Cell viability of CSCs isolated from SUM159PT cells on CAP exposure.

Den för oss kände australiensiske HoloMonitoranvändaren Professor Derek Richards får ett speciellt omnämnande i studien. Det för hans bidrag att visa hur få ut bästa info ur instrumentet.

Acknowledgments

We thank Prof. Derek Richards for helping in the Holomonitor experiments and Dr. Jennifer Gunter for assisting in the metabolism assays.

Resultatet då? Jo,under avsnittet Conclusion skriver forskarna : We demonstrated the synergistic advantages between CAP and Atorvastatin towards enhanced anti-cancer efficacy, and advocated CAP as a promising onco-therapy, functioning alone or as an adjunct to other therapeutic modalities towards the hope of cancer eradication.

Min kommentar

Än en gång visar HoloMonitor sin användbarhet i praktiskt utförande. Med instrumentets hjälp har forskarnas studier visat på en ev ny behandlingsform för de olycksaliga kvinnor som drabbats av den idag svårbotade typen trippelnegativ bröstcancer.

                                              Mvh the99

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