Obtaining Human Breast Adipose Cells for Breast Cancer Cell Co-culture Studies

Primary human breast cancers invade surrounding fat and contact adipocytes, inflammatory infiltrates, and fibrous stroma. This tissue niche influences breast tumor progression. Here, we present a protocol to enable the in vitro study of the complex interactions that occur between breast cancer cells and adipose cells. We describe how to obtain different adipose cell populations, including adipose-derived stem cells, immature adipocytes, and mature adipocytes, from human breast fat tissue and detail the application for co-culture assays with breast cancer cells. For complete details on the use and execution of this protocol, please refer to Picon-Ruiz et al. (2016) and Qureshi et al. (2020).This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 84510

Biofabrication of a tri-layered 3D-bioprinted CSC-based malignant melanoma model for personalized cancer treatment

Conventional in vitro cancer models do not accurately reproduce the tumor microenvironment (TME), so three-dimensional (3D)-bioprinting represents an excellent tool to overcome their limitations. Here, two multicellular tri-layered malignant melanoma (MM) models composed by cancer stem cells (CSCs) isolated from a MM established cell line or a primary-patient derived cell line, fibroblasts, mesenchymal stem cells, and endothelial cells, embedded within an agarose-collagen type I hydrogel were bioprinted. Embedded-cells showed high proliferation and metabolic activity, and actively remodeled their TME. MM hydrogels displayed similar rheological properties that skin and were able to support an early onset of vascularization. Besides, MM hydrogels displayed different response to vemurafenib compared with cell cultures, and supported tumorigenesis in murine xenotransplant achieving more mimetic in vivo models. For the first time a tri-layered 3D-bioprinted CSC-based human MM model is developed recreating TME in vitro and in vivo and response to treatment, being useful for precision treatment regimens against MM.Consejería de Salud y Familias de la Junta de Andalucía (Project No. PIN-0224-2019)Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía (FEDER Funds, Projects PY18-FR-2470, B-CTS-230-UGR18, A-CTS-180-UGR20, PYC20 RE 015 UGR and P18-FR-2465),Ministry de Economía y Competitividad, Instituto de Salud Carlos III (FEDER funds, Projects Nos. DTS19/00145 and DTS21/00098)Chair ‘Doctors Galera-Requena in cancer stem cell research’ (CMCCTS963)Universidad de Granada/CBUA.Plan Andaluz de Investigación, Desarrollo e Innovación (PAIDI 2020— FEDER funds—Ref: DOC_01574)

Lipid-core nanoparticles: Classification, preparation methods, routes of administration and recent advances in cancer treatment

Nanotechnological drug delivery platforms represent a new paradigm for cancer therapeutics as they improve the pharmacokinetic profile and distribution of chemotherapeutic agents over conventional formulations. Among nanoparticles, lipid-based nanoplatforms possessing a lipid core, that is, lipid-core nanoparticles (LCNPs), have gained increasing interest due to lipid properties such as high solubilizing potential, versatility, biocompatibility, and biodegradability. However, due to the wide spectrum of morphologies and types of LCNPs, there is a lack of consensus regarding their terminology and classification. According to the current state-of-the-art in this critical review, LCNPs are defined and classified based on the state of their lipidic components in liquid lipid nanoparticles (LLNs). These include lipid nanoemulsions (LNEs) and lipid nanocapsules (LNCs), solid lipid nanoparticles (SLNs) and nanostructured lipid nanocarriers (NLCs). In addition, we present a comprehensive and comparative description of the methods employed for their preparation, routes of administration and the fundamental role of physicochemical properties of LCNPs for efficient antitumoral drug-delivery application. Market available LCNPs, clinical trials and preclinical in vivo studies of promising LCNPs as potential treatments for different cancer pathologies are summarized.MCIN/AEI FPU18/05336European Social Fund (ESF)Ph.D. program of Biomedicine of the University of GranadaMCIN/AEI/FEDER "Una manera de hacer Europa" RTI2018.101309B-C21 RTI2018.101309B-C2

Clinical implications of inflammation in atheroma formation and novel therapies in cardiovascular diseases

PH-C is supported by an FPU grant from the Ministry of Education, Culture and Sport. This work has been partially funded by the University of Jaen, Acción I apoyo a la investigación (BIO-349) and by Modeling Nature (MNat), Project number QUAL21-11.Cardiovascular diseases (CVD) are the leading causes of death and disability in the world. Among all CVD, the most common is coronary artery disease (CAD). CAD results from the complications promoted by atherosclerosis, which is characterized by the accumulation of atherosclerotic plaques that limit and block the blood flow of the arteries involved in heart oxygenation. Atherosclerotic disease is usually treated by stents implantation and angioplasty, but these surgical interventions also favour thrombosis and restenosis which often lead to device failure. Hence, efficient and long-lasting therapeutic options that are easily accessible to patients are in high demand. Advanced technologies including nanotechnology or vascular tissue engineering may provide promising solutions for CVD. Moreover, advances in the understanding of the biological processes underlying atherosclerosis can lead to a significant improvement in the management of CVD and even to the development of novel efficient drugs. To note, over the last years, the observation that inflammation leads to atherosclerosis has gained interest providing a link between atheroma formation and oncogenesis. Here, we have focused on the description of the available therapy for atherosclerosis, including surgical treatment and experimental treatment, the mechanisms of atheroma formation, and possible novel therapeutic candidates such as the use of anti- inflammatory treatments to reduce CVD.FPU grant from the Ministry of Education, Culture and SportUniversity of Jaen, Accion I apoyo a la investigacion BIO-349Modeling Nature (MNat) QUAL21-1

Dual Role of Fibroblasts Educated by Tumour in Cancer Behavior and Therapeutic Perspectives

Tumours are complex systems with dynamic interactions between tumour cells, nontumour cells, and extracellular components that comprise the tumour microenvironment (TME). The majority of TME’s cells are cancer-associated fibroblasts (CAFs), which are crucial in extracellular matrix (ECM) construction, tumour metabolism, immunology, adaptive chemoresistance, and tumour cell motility. CAF subtypes have been identified based on the expression of protein markers. CAFs may act as promoters or suppressors in tumour cells depending on a variety of factors, including cancer stage. Indeed, CAFs have been shown to promote tumour growth, survival and spread, and secretome changes, but they can also slow tumourigenesis at an early stage through mechanisms that are still poorly understood. Stromal–cancer interactions are governed by a variety of soluble factors that determine the outcome of the tumourigenic process. Cancer cells release factors that enhance the ability of fibroblasts to secrete multiple tumour-promoting chemokines, acting on malignant cells to promote proliferation, migration, and invasion. This crosstalk between CAFs and tumour cells has given new prominence to the stromal cells, from being considered as mere physical support to becoming key players in the tumour process. Here, we focus on the concept of cancer as a non-healing wound and the relevance of chronic inflammation to tumour initiation. In addition, we review CAFs heterogeneous origins and markers together with the potential therapeutic implications of CAFs “re-education” and/or targeting tumour progression inhibition.Consejería de Economía, Conocimiento, Empresas y Universidad de la Junta de Andalucía and European Regional Development Fund (ERDF), ref. P18-FR-2470,Ministry of Science, Innovation and Universities (ref. RTI2018-101309-B-C22)Chair “Doctors Galera-Requena in cancer stem cell research” (CMC-CTS963

Silver Nanoparticles from Annona muricata Peel and Leaf Extracts as a Potential Potent, Biocompatible and Low Cost Antitumor Tool

Cancer is one of the most prevalent diseases in the world and requires new therapies for its treatment. In this context, the biosynthesis of silver nanoparticles (AgNPs) has been developed to treat different types of tumors. The Annona muricata plant is known for having anticancer activity. Its main compounds present in the leaves, stems and skin, allowing for its use as reducing agents. In this manuscript, AgNPs with leaf extract (AgNPs-LE) and fruit peel extract (AgNPs-PE) of A. muricata were biosynthesized obtaining an average nanoparticle diameter sizes smaller than 50 nm, being 19.63 ± 3.7 nm and 16.56 ± 4.1 nm, and with a surface plasmonic resonance (SPR) at 447 and 448 nm, respectively. The lactone functional group present in the LE and PE extracts was identified by the FTIR technique. The behavior and antiproliferation activity of AgNPs-LE and AgNPs-PE were evaluated in breast, colon and melanoma cancer cell lines. Our results showed that Annona muricata fruit peel, which is a waste product, has an antitumor effect more potent than leaf extract. This difference is maintained with AgNPs where the destruction of cancer cells was, for the first time, achieved using concentrations that do not exceed 3 μg/mL with a better therapeutic index in the different tumor strains. In conclusion, we present a low-cost one-step experimental setup to generate AgNPs-PE whose in-vitro biocompatibility and powerful therapeutic effect make it a very attractive tool worth exploiting.Fundacion Empresa Universidad de Granada (Project PR/18/001)Fundación Mutua Madrileña (Project FMM-AP16683-2017)Consejería de Salud Junta de Andalucía (PI-0089-2017)Instituto de Salud Carlos III (RTI2018-101309-B-C22

Unifying Different Cancer Theories in a Unique TumourModel: Chronic Inflammation and Deaminases as Meeting Points

The increase in cancer incidences shows that there is a need to better understand tumour heterogeneity to achieve efficient treatments. Interestingly, there are several common features among almost all types of cancers, with chronic inflammation induction and deaminase dysfunctions singled out. Deaminases are a family of enzymes with nucleotide-editing capacity, which are classified into two main groups: DNA-based and RNA-based. Remarkably, a close relationship between inflammation and the dysregulation of these molecules has been widely documented, which may explain the characteristic intratumor heterogeneity, both at DNA and transcriptional levels. Indeed, heterogeneity in cancer makes it difficult to establish a unique tumour progression model. Currently, there are three main cancer models—stochastic, hierarchic, and dynamic—although there is no consensus on which one better resembles cancer biology because they are usually overly simplified. Here, to accurately explain tumour progression, we propose interactions among chronic inflammation, deaminases dysregulation, intratumor genetic heterogeneity, cancer phenotypic plasticity, and even the previously proposed appearance of cancer stem-like cell populations in the edges of advanced solid tumour masses (instead of being the cells of origin of primary malignancies). The new tumour development model proposed in this study does not contradict previously accepted models and it may open up a window to interesting therapeutic approaches.FPU grant from the Ministry of Education, Culture and SportUniversity of Jaen, Accion I apoyo a la investigacion BIO-349Excellence Research Unit "Modeling Nature" (MNat)Junta de Andalucia European Commission SOMM17/6109/UGRConsejeria de Salud y Familias de la Junta de Andalucia (FEDER funds) PEMP-0205-2020Ministry of Economy and Competitiveness (FEDER funds) PIE16/00045Ministry of Science, Innovation and Universities RTI2018-101309-B-C22Chair "Doctors Galera-Requena in cancer stem cell research" CMC-CTS96

Smart Drug-Delivery Systems for Cancer Nanotherapy

Despite all the advances achieved in the field of tumor-biology research, in most cases conventional therapies including chemotherapy are still the leading choices. The main disadvantage of these treatments, in addition to the low solubility of many antitumor drugs, is their lack of specificity, which explains the frequent occurrence of serious side effects due to nonspecific drug uptake by healthy cells. Progress in nanotechnology and its application in medicine have provided new opportunities and different smart systems. Such systems can improve the intracellular delivery of the drugs due to their multifunctionality and targeting potential. The purpose of this manuscript is to review and analyze the recent progress made in nanotherapy applied to cancer treatment. First, we provide a global overview of cancer and different smart nanoparticles currently used in oncology. Then, we analyze in detail the development of drug-delivery strategies in cancer therapy, focusing mainly on the intravenously administered smart nanoparticles with protein corona to avoid immune-system clearance. Finally, we discuss the challenges, clinical trials, and future directions of the nanoparticle-based therapy in cancer.Comment: Preprint version, 25 pages, 7 figures, 3 tables. Authors thank to Bentham Science the posibility of deposit the ACCEPTED VERSION of the peer-reviewed article after 12 months of publication on journal web site on arXiv repository. The published manuscript is available at EurekaSelect via https://www.eurekaselect.com/openurl/content.php?genre=article&doi=10.2174/138945011766616052714254

Smart drug-delivery systems for cancer nanotherapy

Authors thanks Bentham Sciencew for allow the deposit of the ACCEPTED VERSION of this peer-reviewed article in this institutional repository. Commons License CC BY-NC-ND 4.0 - Attribution-NonCommercial-NoDerivatives 4.0 International. The published manuscript is available at EurekaSelect via https://www.eurekaselect.com/openurl/content.php? genre=article&doi=10.2174/1389450117666160527142544Despite all the advances achieved in the field of tumor-biology research, in most cases conventional therapies including chemotherapy are still the leading choices. The main disadvantage of these treatments, in addition to the low solubility of many antitumor drugs, is their lack of specificity, which leads to the occurrence of severe side effects due to nonspecific drug uptake by healthy cells. Progress in nanotechnology and its application in medicine have provided new opportunities and different smart systems. Such systems can improve the intracellular delivery of the drugs due to their multifunctionality and targeting potential. The purpose of this manuscript is to review and analyze the recent progress made in cancer nanotherapy. First, we provide a global overview of cancer and different smart nanoparticles currently used in oncology. Then, we analyze in detail the development of drug-delivery strategies in cancer therapy, focusing mainly on the intravenously administered smart nanoparticles with protein corona to avoid immune-system clearance. Finally, we discuss the challenges, clinical trials, marketed nanomedicines and future directions of the nanotherapy applied to cancer treatment.This work was supported by the projects MAT2013-43922-R, MAT2015-63644-C2-2-R, (European FEDER support included, MINECO Spain), PI-0533-2014 (Fundación Progreso y Salud/Janssen Cilag, Junta de Andalucía, Spain) and P07-FQM2496, P10-CTS-6270 and P07- FQM3099 (Junta de Andalucía, Spain)

Evolution of Metastasis Study Models toward MetastasisOn-A-Chip: The Ultimate Model?

For decades, several attempts have been made to obtain a mimetic model for the study of metastasis, the reason of most of deaths caused by cancer, in order to solve the unknown phenomena surrounding this disease. To better understand this cellular dissemination process, more realistic models are needed that are capable of faithfully recreating the entire and essential tumor microenvironment (TME). Thus, new tools known as tumor-on-a-chip and metastasis-on-a-chip have been recently proposed. These tools incorporate microfluidic systems and small culture chambers where TME can be faithfully modeled thanks to 3D bioprinting. In this work, a literature review has been developed about the different phases of metastasis, the remaining unknowns and the use of new models to study this disease. The aim is to provide a global vision of the current panorama and the great potential that these systems have for in vitro translational research on the molecular basis of the pathology. In addition, these models will allow progress toward a personalized medicine, generating chips from patient samples that mimic the original tumor and the metastatic process to perform a precise pharmacological screening by establishing the most appropriate treatment protocol.Ministerio de Educacion y Formacion Profesional FPU19/02138 BOE-B-2019-44783Ph.D. program of Biomedicine of the University of GranadaConsejeria de Economia, Conocimiento, Empresas y Universidad, Junta de Andalucia (FEDER Funds) P18-FR-2470 B-CTS-230-UGR18 SOMM17/6109/UGRInstituto de Salud Carlos III (FEDER Funds) PIE16-00045 DTS19/00145Junta de Andalucia PIN-0224-2019Chair "Doctors Galera-Requena in cancer stem cell research" CMC-CTS96