Applying the Free-Energy Principle to Complex Adaptive Systems

The free energy principle is a mathematical theory of the behaviour of self-organising systems that originally gained prominence as a unified model of the brain. Since then, the theory has been applied to a plethora of biological phenomena, extending from single-celled and multicellular organisms through to niche construction and human culture, and even the emergence of life itself. The free energy principle tells us that perception and action operate synergistically to minimize an organism’s exposure to surprising biological states, which are more likely to lead to decay. A key corollary of this hypothesis is active inference—the idea that all behavior involves the selective sampling of sensory data so that we experience what we expect to (in order to avoid surprises). Simply put, we act upon the world to fulfill our expectations. It is now widely recognized that the implications of the free energy principle for our understanding of the human mind and behavior are far-reaching and profound. To date, however, its capacity to extend beyond our brain—to more generally explain living and other complex adaptive systems—has only just begun to be explored. The aim of this collection is to showcase the breadth of the free energy principle as a unified theory of complex adaptive systems—conscious, social, living, or not

A Novel 3D In Vitro Platform for Pre-Clinical Investigations in Drug Testing, Gene Therapy, and Immuno-oncology.

Tumors develop within complex cell-to-cell interactions, with accessory cells playing a relevant role starting in the early phases of cancer progression. This event occurs in a three-dimensional (3D) environment, which to date, has been difficult to reproduce in vitro due to its complexity. While bi-dimensional cultures have generated substantial data, there is a progressive awareness that 3D culture strategies may rapidly increase the understanding of tumor development and be used in anti-cancer compound screening and for predicting response to new drugs utilizing personalized approaches. However, simple systems capable of rapidly rebuilding cancer tissues ex-vivo in 3D are needed and could be used for a variety of applications. Therefore, we developed a flat, handheld and versatile 3D cell culture bioreactor that can be loaded with tumor and/or normal cells in combination which can be monitored using a variety of read-outs. This biocompatible device sustained 3D growth of tumor cell lines representative of various cancers, such as pancreatic and breast adenocarcinoma, sarcoma, and glioblastoma. The cells repopulated the thin matrix which was completely separated from the outer space by two gas-permeable membranes and was monitored in real-time using both microscopy and luminometry, even after transportation. The device was tested in 3D cytotoxicity assays to investigate the anti-cancer potential of chemotherapy, biologic agents, and cell-based therapy in co-cultures. The addition of luciferase in target cancer cells is suitable for comparative studies that may also involve parallel in vivo investigations. Notably, the system was challenged using primary tumor cells harvested from lung cancer patients as an innovative predictive functional assay for cancer responsiveness to checkpoint inhibitors, such as nivolumab. This bioreactor has several novel features in the 3D-culture field of research, representing a valid tool useful for cancer investigations, drug screenings, and other toxicology approaches

Current treatments for non-small cell lung carcinoma

This literature review addresses the subject of non-small-cell lung carcinoma (NSCLC) and the current treatments for it. For therapeutic purposes, lung carcinomas are histologically grossly organized into two main groups that are NSCLC and small-cell lung carcinoma (SCLC). NSCLC comprises about 85% of all lung carcinoma cases and are further organized into three histologic subgroups: squamous-cell carcinoma, adenocarcinoma, and large-cell lung cancer. Smoking status and mortality rates have been widely documented to have a correlation with all lung cancers. Diagnostical tools have improved and are sufficient to establish an accurate diagnosis without the need to rely on immunocytochemical or immunohistochemical analysis on routine basis. Despite these advanced tools and benefits of receiving early intervention, NSCLC is often di agnosed late, which precedes poor prognosis. Lung cancer develops over long period of time and often presents itself with non-specific res piratory symptoms, such as coughing, shortness of breath and sometimes bloody sputum. With smokers already excessively represented in pulmonary diseases and simultaneously being the dominant bracket to develop NSCLC, some of the warning signs of this cancer might go unnoticed for longer period amongst them. Current treatment options include traditional cancer treatments such as surgery, radiotherapy, and chemotherapy. Emphasis on these has remained strong, but treatments have begun convert into new and more specific options. Combination therapies, targeted therapies and immunotherapies are increasingly used in late-stage disease. Lastly, we touch on the possibil ities of novel bacteria-based therapies for lung cancer. Keywords: non-small-cell lung cancer, surgery, radiotherapy, chemotherapy, immunother apy, targeted therapy, bacteria-based therap

Array-CGH in the investigation of karyotype changes of CD34+ haematopoietic stem cells in lymphoma and multiple myeloma patients who underwent to autologous transplantation.

openHematopoietic stem cell transplantation (HSCT) represents an effective treatment strategy for a variety of hematologic and not hematologic malignancies. In particular, autologous transplantation of haematopoietic stem cells (ASCT) from bone marrow of patients with hematologic malignancies is feasible and has low treatment-related mortality (Gribben JG, 2009). However, literature assessed late mortality in 29.4% of individuals who had survived 2 or more years after autologous haematopoietic cell transplantation (Burns, L.J., 2009; Bhatia S. and al, 2005). This could be due to the purified stem cells which might carry a mutation on a chromosome predisposing to the disease and lead to the risk of pathology recurrence. Lymphoma and myeloma are perfect candidates for autologous transplantation after G-CSF stimulation and bone marrow ablation trough chemotherapy. However, in this case too, pathology relapse or secondary malignancies are found in a high percentage of patients. The aim of this project is to verify the existence of detectable imbalanced chromosome anomalies in stem cells before any ablative treatment for HSCT or developed after G-CSF stimulation or chemotherapy. A cohort of 24 lymphoma and myeloma patients have been analyzed trough array-CGH to identify significant imbalanced chromosome anomalies also present in low percentage of mosaicism. The result showed anomalies in 8/24 patients: one patient affected by Hodgkin Lymphoma (HL) revealed a deletion of chromosome 2 in p16.1, where the REL gene is located and in part deleted; the amplification of chromosome 11 in q12.2q13.4 containing CCND1 gene (this patient was investigated both before and after transplantation) was found in one patient with multiple myeloma (MM); alterations of chromosome 14 in q32.31-33, where genes for variable chain of immune globulin are located, were found in five patients with Hodgkin and non Hodgkin lymphomas (HL/NHL). FISH on interphase nuclei has been used to confirm a-CGH data. A short-time (36 months) clinical and haematological follow-up examination did not show a different trend between patients with chromosome imbalances and without but a long-term follow-up is needed to definitely correlate the imbalances with the clinical evolution and to have the indications of global survival of the considered population. Work in progress is the extension of clinical and haematological observation to obtain evidence of a difference statistically significant and to reach the final goal of suggesting a possible protocol to candidate patients to purging treatments before the CD34+ cells re-infusion.openMedicina Sperimentale e OncologiaMare, LydiaMare, Lydi

CIrculating TUmor CELls (CTCs), circulating tumor DNA (ctDNA) and exosomes (Ex) in breast cancer patients: a prospective study

Background The translational study CITUCEL aims to correlate the presence of CTCs and ctDNA at different time points with the clinical outcome of BC patients in neo-adjuvant, adjuvant or advanced setting and to compare the mutational profiles of the primary tumor, CTCs and ctDNA in the different settings. Methods CTCs have been characterized and isolated single or in pool with the DEPArray system, after immunologic negative enrichment with antibodies coupled to magnetic beads against common leukocytes and red blood cells antigens, followed by cells labelling with specific antibody for epithelial, mesenchymal and pan-leukocyte markers. CTCs mutational analyses have been performed by NGS on the Ion Torrent\u2122 platform using the Oncomine Breast cfDNA V2.0 Kit and an in house developed approach based on the molecular tagging. The same kit and the same panel was used to analyse ctDNA, extracted from plasma using ccfDNA LV kit (Promega). Primary tumor mutational profile has been evaluated in NGS with a custom panel. Results A total of 60 patients have been enrolled in the study (32 in neo-adjuvant, 22 in adjuvant and 6 in advanced settings). CTCs have been recovered and morphologically/phenotypically re-evaluated; ctDNA has been extracted, quantified and analysed by NGS. Preliminary analysis shown a significative difference in CTCs baseline counts between the early settings and the advanced one (P=0.038). No significative change has been detected before and after treatment in the (neo)-adiuvant setting, but remarcably a lower CTCs baseline count correlates with pCR (P=0.0399). Similar results have been obtained in regards of ctDNA, definitely higher in the blood of MTS patients (P=0.0127); no significative differences were detected before and after treatment, and no correlation has been found with pCR. The mutational analysis is in progress. Conclusions The results of the study will provide information on the applicability of liquid biopsy in clinical practice

Optimization of the Nutritional Environment for Differentiation of Human Induced Pluripotent Stem Cells using Design of Experiments – A Proof of Concept

The utilization of human-induced pluripotent stem cells (hiPSCs) in cell therapy has a tremendous potential but faces many practical challenges, including costs associated with cell culture media and growth factors. There is an immediate need to establish an optimized culture platform to direct the differentiation of hiPSCs into germ layers in a defined nutritional microenvironment to generate cost-effective and robust therapeutics. The aim of this study was to identify the optimal nutritional environment by mimicking the in vivo concentrations of three key factors (glucose, pyruvate, and oxygen) during the spontaneous differentiation of hiPSCs derived from cord blood, which greatly differ from the in vitro expansion and differentiation scenarios. Moreover, we hypothesized that the high glucose, pyruvate, and oxygen concentrations found in typical growth media could inhibit the differentiation of certain lineages. A design of experiments was used to investigate the interaction between these three variables during the spontaneous differentiation of hiPSCs. We found that lower oxygen and glucose concentrations enhance the expression of mesodermal (Brachyury, KIF1A) and ectodermal (Nestin, β-Tubulin) markers. Our findings present a novel approach for efficient directed differentiation of hiPSCs through the manipulation of media components while simultaneously avoiding the usage of growth factors thus reducing costs