It is becoming increasingly clear that short and long noncoding RNAs critically participate in the regulation of cell growth, differentiation, and (mis)function. However, while the functional characterization of short non-coding RNAs has been reaching maturity, there is still a paucity of well characterized long noncoding RNAs, even though large studies in recent years are rapidly increasing the number of annotated ones. The long noncoding RNA PVT1 is encoded by a gene that has been long known since it resides in the well-known cancer risk region 8q24. However, a couple of accidental concurrent conditions have slowed down the study of this gene, that is, a preconception on the primacy of the protein-coding over noncoding RNAs and the prevalent interest in its neighbor MYC oncogene. Recent studies have brought PVT1 under the spotlight suggesting interesting models of functioning, such as competing endogenous RNA activity and regulation of protein stability of important oncogenes, primarily of the MYC oncogene. Despite some advancements in modelling the PVT1 role in cancer, there are many questions that remain unanswered concerning the precise molecular mechanisms underlying its functioning. Evidence strongly supports pervasive transcription of human genomes, even though only 2% of transcripts encode proteins –5. NoncoNoncoding transcripts are usually classified on the basis of their size : miRNAs  cis and in trans. Many functional lncRNAs have been shown to play key roles in organ development and cancer; however the biological functions of the majority of lncRNAs largely remain to be identified. Increasing experimental evidence supports the hypothesis that lncRNAs may possess competing endogenous RNA (ceRNA) activity [While miRNAs have been extensively studied –10, lncRactivity , 17–21. Our group recently developed a computational model able to identify putative ceRNAs among lncRNAs  and usedThe human PVT1 gene  is a long intergenic noncoding RNA (lincRNA) homologous to the mouse plasmacytoma variant translocation gene (Pvt1). Indeed this locus has been first discovered in the mid 80s in mouse as frequently involved in a variant translocation in plasmacytomas , 24. ShoIn the light of all the above, we can hazard a guess that if PVT1 was a protein-coding gene it would have been at the heart of countless studies aimed at the functional characterization since its discovery. Rather, being a noncoding transcript has slowed the process of shading light on its mechanisms of action, for reasons of both technical and cultural nature. In the early years of its discovery, in fact, on the one hand the classical molecular techniques for functional investigation were not suited to detecting and analyzing noncoding RNAs; on the other hand it was hardly conceivable that what was still deemed as transcriptional noise could play an active role in the regulation of important processes.Instead, the technological revolution of recent years along with the unprecedented appreciation of the multifaceted world of noncoding RNAs has led to a renewed interest in the investigation of the PVT1 function. This growing interest is reflected in the multitude of papers published in the last few years on PVT1 . As a co gene desert, so termed due to paucity of transcriptional activity [In humans the chromosomal locus encoding PVT1  maps to activity , 58, whiactivity , 59, 60.activity .Another remarkable feature of the PVT1 locus is to reside some 60 kilobases (kb) 3-prime of the well-known MYC oncogene and different functional interactions between these two genes have been proposed , 62. AddBoth the mouse and the rat homologous of human PVT1  constitute sites of frequent retroviral integration , 63–66 ar = 0.5 and P values = 10−32, The PVT1 gene  spans acBy analysing the breast cancer data set provided by TCGA, we found  that PVTIn terms of topological properties, PVT1 switches from being the first of the hubs in the normal-MMI-network to fall outside the list of nodes of the cancer network. Interestingly, recent studies suggested a role for PVT1 in the pathophysiology of breast cancer by virtue of PVT1-mediated inhibition of apoptosis and increasing of proliferation, when overexpressed .Among competitors of PVT1 for binding to the mir-200 family in normal breast tissues, there are some important regulators of breast tissue morphogenesis and development, such as CDH1; all the three members of the extended p53 family ; two members of the mammalian RUNX family ; and GATA3. CDH1  is a crucial cell-cell adhesion molecule considered as an epithelial cell marker. Loss of function of CDH1 has been associated with increased proliferation, invasiveness, and/or metastasis in several epithelial cancers, including breast cancer . TP53 isThus, the PVT1 neighborhood in the normal-MMI-network encompasses cancer related genes as well as genes involved in mammary gland development and cell morphogenesis and the sponge program orchestrated by PVT1 results completely abolished in cancer. Taken together these findings may be indicative of a possible PVT1 surveillance role aimed to preserve cell-cell adhesion. Indeed, mammary gland morphogenesis results from the coordination of diverse cellular processes involving cell-cell adhesion, migration, proliferation, and apoptosis. Thus, PVT1 controlling circuit may provide further insight in solving this complicated puzzle.The specific conditions required for ceRNA network to occur are still far from being determined. The importance of the relative concentration of the ceRNAs, as well as their related miRNAs, has been recently emphasized . Large cA completely different way of functioning of PVT1 comes from new findings that highlight its involvement in the regulation of the oncoprotein MYC . Accordiβ1 signaling pathway  lncRNAs in a mouse model for hepatocellular carcinoma (HCC) supported the role for PVT1 in regulating proliferation and the same phenotype was confirmed in human HCC cell lines . In line pathway  and sugghway (β1 . Intereshway (β1 . Remarkahway (β1 .A wide range of solid tumors often carry amplification of the 8q24 locus, a large genomic region spanning the PVT1 gene and its bulky neighboring MYC. Despite early recognition of the importance of this locus with respect to many malignancies, the well-known MYC oncogene has repeatedly been considered the first guilt by association. While the role of MYC in cancer has been proved beyond any doubt, the presence of this famous neighbor together with the enigmatic noncoding nature of PVT1 has hampered prompt recognition that another important oncogene coexisted with MYC in the 8q24 risk region. Nowadays, the accumulating evidences showing that PVT1 and MYC have separate and distinct oncogenic potential , 32, 86 In conclusion, the foregoing considerations suggest to think that the time has come for a thorough investigation of the role played by PVT1 in tumors, a challenge full of promise.


Mitochondrion is known as the energy factory of the cell, which is also a unique mammalian organelle and considered to be evolved from aerobic prokaryotes more than a billion years ago. Mitochondrial DNA, similar to that of its bacterial ancestor’s, consists of a circular loop and contains significant number of unmethylated DNA as CpG islands. The innate immune system plays an important role in the mammalian immune response. Recent research has demonstrated that mitochondrial DNA (mtDNA) activates several innate immune pathways involving TLR9, NLRP3 and STING signaling, which contributes to the signaling platforms and results in effector responses. In addition to facilitating antibacterial immunity and regulating antiviral signaling, mounting evidence suggests that mtDNA contributes to inflammatory diseases following cellular damage and stress. Therefore, in addition to its well-appreciated roles in cellular metabolism and energy production, mtDNA appears to function as a key member in the innate immune system. Here, we highlight the emerging roles of mtDNA in innate immunity. Microorganisms that cause disease in humans and animals enter the body at different sites and produce disease symptoms via different mechanisms. Innate immune response, or innate immunity, could immediately act as the first line of defense against various pathogens and do not involve the activation of antigen-specific lymphocytes. Innate immunity is sufficient to prevent the body from being routinely overwhelmed by a wide range of microorganisms that live on or in it Kenneth, .Mitochondria are known as the energy factories of the cell, which are also unique mammalian organelles and considered to be evolved from aerobic prokaryotes more than a billion years ago  in inflammation initiation through Toll-like receptor 9 (TLR9) was characterized and its key role in provoking the NLRP3 inflammasome was also reported. Mitochondrial DNA also triggers stimulator of interferon genes (STING) signaling.In recent years, the role of mtDNA as DAMPs is recognized, which are endogenous molecules released by cells undergoing abnormal cell death  and capable of activating innate immune response. DAMPs could be recognized by same receptors in pathogen-associated molecular patterns (PAMPs) recognition, such as the pattern recognition receptors (PRRs). Recently, the role of mtDNA as DAMPs in inflammation initiation has gained much attention.in vivo, which is mediated by a pathway involving TLR9 signaling. The injection of mtDNA can activate neutrophils and increase the release of matrix metalloproteinase-8 (MMP-8), a pro-inflammatory cytokine, leading to severe inflammation in mouse lungs. Moreover, the activation of neutrophils by mtDNA through TLR9 pathway was confirmed by using TLR9−/− mice and TLR9 antagonist ODN2088, and the inflammation was reduced in both TLR9−/−and ODN2088-treated mice  family, pyrin domain containing 3 (NLRP3) inflammasome is the most extensively investigated one among the identified inflammasomes, which is mainly due to its ability to be activated by a wide variety of ligands  patients but not in healthy individuals  based on sensitivity to cyclosporine A, which is plausible if MPT is followed by osmotic swelling and mitochondrial rupture (Nakahira et al.,


Ischemic stroke is characterized by a rapid loss of brain function due to disturbance in blood supply to a part of the brain. Due to fixed intracranial space, any increase in intracranial fluid volume, or progressive brain oedema formation, contributes to further deterioration of the already impaired brain function. Bradykinin (BK), which levels increase during ischemic stroke, promotes blood–brain barrier permeability and raises intracranial capillary blood pressure, leading to brain oedema formation. Furthermore, BK induces glutamate release from neurons and astrocytes via activation of BK receptor type 2. suggesting involvement of BK in glutamate neurotoxicity. It has been recently shown that humans without functional natriuretic peptides (NPs) suffer from massive stokes ,2.NPs can reduce brain oedema and have a neuroprotective role in acute ischemic stroke as well as during recovery after stroke. Although mechanisms are still not clear, it appears that NPs enhance angiogenesis, neurogenesis and oligodenrogenesis ,4. One o2+ concentration.Aim of our study is to determine beneficial effects of the NPs in stroke development in murine model . The symptoms of the stroke are determined by behavioural studies. The sizes of the lesion and brain oedema are established by μCT. Furthermore, we determined the effects of NPs on the BK signalling pathway in primary culture of neurons and astrocytes using whole cell patch clamp experiments to measure membrane potential and measurements of intracellular Ca2+ concentration of astrocytes and neurons, followed by activation of Ca2+-dependent Cl- channel which depolarized the cell membrane. Agonists of guanylate cyclase A, partially guanylate cyclase C but not guanylate cyclase B inhibited the effects of BK at the membrane potential and intracellular Ca2+ concentration via regulators of G protein signalling. In vivo experiments showed that urodilatin inhibited development of stroke symptoms, the formation of the ischemic lesion and brain oedema.In primary isolated astrocytes and neurons, BK binding to type 2 receptor, leads to an increase in intracellular CThe results of this research show the existence of a natural antagonist of the BK receptor type 2 in the mouse brain, and the possible use of NPs in treatment of the stroke.


Streptozotocin (STZ), a glucosamine-nitrosourea compound derived from soil bacteria and originally developed as an anticancer agent, in 1963 has been found to induce diabetes in experimental animals. Since then, systemic application of STZ became the most frequently studied experimental model of insulin-dependent (type 1) diabetes. The compound is selectively toxic toward insulin-producing pancreatic beta cells, which is explained as the result of its cellular uptake by the low-affinity glucose transporter 2 (GLUT2) protein located in their cell membranes. STZ cytotoxicity is mainly due to DNA alkylation which results in cellular necrosis. Besides pancreatic beta cells, STZ applied systemically damages also other organs expressing GLUT2, such as kidney and liver, whereas brain is not affected directly because blood-brain barrier lacks this transporter protein. However, single or double intracerebroventricular (icv) STZ injection(s) chronically decrease cerebral glucose uptake and produce multiple other effects that resemble molecular, pathological, and behavioral features of Alzheimer’s disease (AD). Taking into consideration that glucose hypometabolism is an early and persistent sign of AD and that Alzheimer’s brains present features of impaired insulin signaling, icv STZ injections are exploited by some investigators as a non-transgenic model of this disease and used for preclinical testing of pharmacological therapies for AD. While it has been assumed that icv STZ produces cerebral glucose hypometabolism and other effects directly through desensitizing brain insulin receptors, the evidence for such mechanism is poor. On the other hand, early data on insulin immunoreactivity showed intense insulin expression in the rodent brain, and the possibility of local production of insulin in the mammalian brain has never been conclusively excluded. Also, there are GLUT2-expressing cells in the brain, in particular in the circumventricular organs and hypothalamus; some of these cells may be involved in glucose sensing. Thus, icv STZ may damage brain glucose insulin producing cells and/or brain glucose sensors. Mechanistic explanation of the mode of action of icv STZ, which is currently lacking, would provide a valuable contribution to the field of animal models of Alzheimer’s disease. Laboratory animals, mostly rodents, are commonly used as models of human diseases. The concept of such models is based on a certain degree of “analogy” between pathological processes that occur during progression of a disease in patients and in animals being studied as their substitute . AlthougThe case of Alzheimer’s disease (AD) is no exception. During the first half of the 1990 decade, mutations of the three “Alzheimer’s genes” coding beta-amyloid precursor protein (β-APP) and presenilins PS-1, PS-2) have been determined as causative in the early onset autosomal dominant forms of this disease   have bee. Since tWhile transgenic models are still the mainstream of AD research, alternative models are sought, which could prove more useful for translational research. The expert advisory panel gathered under the auspice of the Alzheimer’s Drug Discovery Foundation in the report published in 2011  concludeAfter the discovery of the Alzheimer’s genes, the leading concept underlying research in the AD area has been the “amyloid cascade” hypothesis proposed more than 20 years ago . Its ori18FDG-PET technique in individuals harboring Alzheimer’s genes have shown that signs of depressed brain glucose uptake, apparently independent of structural atrophy, are detectable up to three decades prior to the expected onset of clinical symptoms of early onset familial AD 695 expretabolism  followin spectra . Proamyl15O2), glucose consumption (with 19 F-deoxyglucose) and amyloid β burden (with 13C-PIB) and found that the spatial distribution of aerobic glycolysis in normal young adults correlated with the location of Aβ deposits in AD patients as well, as in cognitively normal participants with elevated Aβ. The authors suggested a possible link between regional aerobic glycolysis in young adulthood and later development of Alzheimer pathology.In this context, it may be worth to mention that correlations have been found between deposition of amyloid plaques and the set of frontal and parietal midline structures with high connectivity and high metabolic activity , experimental paradigm as a model of Alzheimer’s disease will be preceeded with a brief summary of the history and properties of this substance. Streptozotocin , single intravenous injection of STZ results in dose-dependent diabetogenic response consisting of increase in serum glucose level and urine volume and decrease in serum and pancreatic immunoreactive insulin; doses of 45–65 mg/kg decreased insulin by more than 10 times . Current+ and ultimately depletion of ATP stores. Besides insulin-producing pancreatic beta cells, STZ is toxic toward other organs expressing GLUT2 transporter, particularly kidney and liver. Brain is not affected directly because STZ is not able to penetrate blood-brain barrier which lacks GLUT2 transporter.Current understanding of the diabetogenic property of streptozotocin and mechanisms involved has been summarized by Lenzen , 33. UnlO-methyl-glucose. However, mechanisms of their diabetogenic activity seem to differ in some points. One difference is that D-glucose and D-mannose are reportedly able to block toxic effect of alloxan but not STZ [Streptozotocin action is considered similar to that of the other well-known diabetogenic substance alloxan. Both compounds are taken up by GLUT2 transporters, act intracellularly, are selectively toxic toward pancreatic insulin-producing β cells, and their toxicity can be blocked by peripheral application of non-metabolysable glucose analog 3- not STZ . The oth not STZ , althoug not STZ .Cytotoxicity of STZ is dependent on the activity of poly (ADP-ribose) polymerase (PARP), and PARP-knockout mice are resistant to this toxin , 36. CurGLUT2 glucose transporting protein is a part of the distributed glucose sensing mechanism which is located in the pancreatic beta cells and some other organs, e.g., liver . GLUT2 iIntracerebroventricular instillations of drugs are rarely used in the clinic for administering drugs ; more frMore than 30 years ago, icv injections of alloxan were used, apparently for the first time, in experiments aimed at elucidating intracerebral glucose sensing and its physiological role . InteresLacković and Salković  comparedThe pioneer of the icv STZ model is Professor Sigfried Hoyer. His early works on this subject have been summarized in the paper published in 1994 . CurrentThe starting point of the reasoning developed by Hoyer was that in the sporadic Alzheimer’s disease, there is a fall in brain consumption of both oxygen and glucose, but a fall in brain oxygen consumption is disproportionately slighter. These observations led to the hypothesis that the major biochemical perturbation in incipient sporadic AD concerns the control over cerebral glucose metabolism, which is subsequent to a signal transduction failure of the cerebral insulin receptor. At this point, another observation was also taken into account, namely that in adult male Wistar rats, intracerebroventricular administration of 80 mU insulin stimulated two key glycolytic enzymes, hexokinase and phosphofructokinase, by approx. 20 %. This later result was interpreted as the evidence for the control of glycolytic flux in the brain by insulin and led to the conclusion that any perturbation in insulin signal transduction will have severe impact on brain glucose metabolism . The nexBased on the close analogy between human patients with AD and rats subjected to icv STZ injections in respect to brain metabolic and behavioral disturbances, Hoyer proposed that icv STZ shall be recognized as the non-transgenic preclinical model of Alzheimer’s disease. In the most recent review , this opDevelopment of several Alzheimer-like indices of brain metabolic and behavioral disturbances in rats subjected to one or two icv STZ injection(s) is considered to be a dynamic process. In the majority of papers, the effects of icv STZ on brain function and behavior were followed within a relatively short time of a few weeks. Only recently, the Hoyer’s group reported on the longer follow-up of icv STZ rats which revealed deposits of amyloid β in the wall of meningeal and cortical blood vessels visualized by thioflavine-S and Congo red as well, as by immunohistochemical staining . In one The other authors also reported on various neurochemical and neuropathological consequences of icv STZ injection(s). Most observations were convergent with the Hoyer’s observations that metabolic disturbances following icv STZ cover large areas of the brain, including locations distant from the injection site, and that they mount up in time. For example, two daily injections of STZ into the lateral ventricles of adult rats caused in the frontoparietal cortex a significant enlargement of the neuronal Golgi apparatus detected 3 weeks later , and simO-GlcNAcylation, as well as increased phosphorylation of tau and neurofilaments and decreased microtubule-binding activity of tau. These authors suggested that the key mechanism in the development of disease in the human AD, as well as in the rodent model disease is the development of brain insulin resistance which leads to neurofibrillary degeneration via two cooperating pathways, namely decreased PI3K-AKT signaling activity and glycogen synthase kinase 3 beta (GSK3β) overactivation and decreased GLUT1/3 expression and decreased intraneuronal glucose metabolism. More recently, investigators from the same laboratory [In the brains of adult icv STZ-treated rats, Deng et al.  found imboratory  comparedboratory  observedRecently, de la Monte  and de lHowever, the results obtained by some other authors are suggestive of spatial restriction of intracerebral damage caused by STZ injection into lateral ventricles. For example, Terwel et al.  injectedIt is frequently assumed that streptozotocin injected intraventricularly damages or desensitizes brain insulin receptors or otherwise produces brain insulin resistance defined as inadequate response of insulin receptors located in the brain to insulin. This interpretation is sometimes presented as an analogy between the mechanism of action of icv STZ on the brain and the mechanism of peripheral action of low doses of STZ. For example, in the  authoritative review on the icv STZ model of AD, Salkovic-Petrisic et al.  opined tIt shall also be taken into account that insulin receptors, at least in some extracranial locations, are subject to intracellular turnover, being continuously synthesized, deposited in cell membranes, endocytosed, and degraded with a relatively short half-life of hours rather than days . If simOf note is also that brain neuron-specific insulin receptor knockout (NIRKO) mice, which exhibit a complete loss of insulin-mediated activation of phosphatidylinositol 3-kinase, were reported to display markedly reduced phosphorylation of Akt and GSK3 beta kinases leading to substantially increased phosphorylation of Tau protein but exhibited no alteration in neuronal proliferation/survival, memory, or basal brain glucose metabolism .The next relevant issue is that of the diffusion of streptozotocin after intracerebrovientricular injection. As pointed out by Pardridge , drug inThe other issue which deserves reflection in the context of icv STZ model of AD is the possibility of intracerebral synthesis of insulin in mammals. In a classical study of Havrankova et al. , no chanInvolvement of GLUT2 as mediator of icv STZ effects has been postulated , 94, 95.N-nitrosodiethylamine (NDEA), the effects were similar [Almost 30 years ago, Coimbra and Migliorini  discover similar .In summary, there is no doubt that single or double injection of streptozotocin into the lateral ventricles of rat produces chronic metabolic, neuropathological, and behavioral disturbances reminiscent of human Alzheimer’s disease, but acceptable mechanistic explanation for these phenomena is lacking. The concept of irreversible inhibition or desensitization of insulin receptors caused directly by STZ diffusing throughout the brain parenchyma does not seem plausible, and alternative explanation(s) shall be sought. Understanding causality between multitude of interacting processes involved in the development of the model disease evoked by icv STZ injection(s) is necessary for assessment of its convergence with the human sporadic Alzheimer’s disease and possible usefulness in translational research.


The signal transducer and activator of transcription (STAT)3 governs essential functions of epithelial and hematopoietic cells that are often dysregulated in cancer. While the role for STAT3 in promoting the progression of many solid and hematopoietic malignancies is well established, this review will focus on the importance of STAT3 in prostate cancer progression to the incurable metastatic castration-resistant prostate cancer (mCRPC). Indeed, STAT3 integrates different signaling pathways involved in the reactivation of androgen receptor pathway, stem like cells and the epithelial to mesenchymal transition that drive progression to mCRPC. As equally important, STAT3 regulates interactions between tumor cells and the microenvironment as well as immune cell activation. This makes it a major factor in facilitating prostate cancer escape from detection of the immune response, promoting an immunosuppressive environment that allows growth and metastasis. Based on the multifaceted nature of STAT3 signaling in the progression to mCRPC, the promise of STAT3 as a therapeutic target to prevent prostate cancer progression and the variety of STAT3 inhibitors used in cancer therapies is discussed. The signal transducer and activator of transcription (STAT)-3 plays an indispensable role in the progression of a wide variety of cancers. Activation of STAT3 downstream of cell surface receptors for cytokines and growth factors, by oncogenes or by chemical carcinogens in the microenvironment of many tumor types drives their transformation, survival, proliferation, invasion and dissemination. This is mediated by phosphorylation of latent cytoplasmic STAT3 on specific residues  by a variety of tyrosine and serine kinases leading to its dimerization and nuclear translocation, where it acts as a transcription factor for a plethora of genes governing the malignant properties of the tumor cell. STAT3 is a critical mediator of differentiation, activation, migration and inflammatory capacity of immune cells and stromal cells that create the microenvironment supporting tumor cell growth . ImportaOverviews on the importance of STAT3 signaling and targeting it with potential therapies for cancer in general have been the subject of recent reviews . Howeveretc.) and Abl etc.) and and Abl , PKC [11 and Abl  is requi and Abl . Once in and Abl . Interes and Abl . vs. IL-10 gene transcription; the transient nature of IL-6 signaling is controlled by rapid SOCS3 inhibition of the gp130 subunit of the IL-6 receptor. This limits the duration that the STAT3 dimer is functional within the nucleus, allowing pro-inflammatory genes with multiple response elements to be transcribed. By contrast, the lack of a SOCS3 binding site in the IL-10 receptor prolongs STAT3 activation, allowing more time for STAT3 dimers to bind to anti-inflammatory genes that have only single STAT3 response elements [et al. found that unphosphorylated-STAT3, through direct interaction with p65 of nuclear factor (NF)-κβ, serves as a suppressor and inhibits the ability of NF-κβ to induce the iNOS promoter in mesangial cells [Importantly, the specificity, duration and inhibition of STAT3 dependent gene transcription is tightly regulated. Because STAT3 signals downstream of so many extracellular cytokines and growth factors and it is activated by so many intracellular kinases, multiple levels of control dictate the specificity of which genes are transcribed in a STAT3 dependent manner, depending on the stimulus. For example, multiple co-activators like CBPp300 , APE1 1 and NCOAelements . In caseelements ,18. For al cells .Growth and maintenance of normal prostate epithelium and primary prostate cancer tumors is fueled by androgen activation of the AR present in prostate cells. This reliance on AR signaling makes androgen depravation therapy (ADT) an effective way to limit prostate cancer tumor growth; initially tumors respond well to anti-androgen therapy, dramatically shrinking in size and patients see a reduction in cancer symptoms. However, tumor recurrence within 3 years of ADT occurs in 80% of patients, and tumor regrowth combined with a rise in the AR dependent production of circulating prostate specific antigen (PSA), marks the progression to CRPC. Continued dependence on AR signaling is a hallmark of CRPC and there are multiple mechanisms by which the AR can be re-activated in castration conditions, including hyperactivation of oncogenic signaling pathways, intratumoral production of androgen, ligand independent AR activation due to AR mutation or splice variants and increased expression of molecular chaperones and the AR itself . Any comPTEN), which is a negative regulator of the PIK3/Akt survival pathway [Hyper-activation of multiple oncogenic signaling pathways drives progression of PCa to CRPC. For example, the most frequently deleted tumor suppressor gene in prostate cancer is phosphatase and tensin homolog gene  to activate the MAPK pathway with short-term IL-6 treatment and promote AR activity [External signals from cytokines and growth factors can trigger oncogenic signaling cascades that modulate AR activity. In particular, IL-6, the prototypical JAK/STAT3 signaling cytokine, can modulate activity and expression of the AR. It is no surprise therefore that increased IL-6 levels in PCa patients has been associated with decreased survival , increasactivity . This inThe link between IL-6 and STAT3 signaling to AR activity is further underscored by the importance of this signaling pathway in the development of anti-androgen resistance. Autocrine IL-6 signaling in PCa cells induces the constitutive activation of STAT3, enhancing recruitment of the AR to the PSA promoter and rendering these cells resistant to the new generation anti-androgen Enzalutamide, which can be reversed by treatment with a STAT3 inhibitor . In addiWhile re-activation of the AR is undoubtedly the hallmark of progression to CRPC, it is also clear that the ability of PCa tumor cells to retain some degree of phenotypic plasticity, or the ability to change into a variety of aggressive tumor cell types, such as cancer stem cells or cells that have undergone epithelial to mesenchymal transition (EMT), is also a critical component of advanced disease. CSC theory postulates that transformed progenitor cells drive continued expansion of tumors with invasive and metastatic propensity . CSCs hain vivo [Emerging evidence suggests that STAT3 plays critical roles in maintaining CSCs and promoting EMT in PCa as well as head and neck, hepatocellular carcinoma, glioblastoma and breast cancers. This is not surprising as a large body of work has shown the importance of STAT3 in embryonic stem cell differentiation in cooperation with other central regulators of pluripotency, including Oct4 and Sox family transcription factors as well as LIF . Importain vivo . The requirement for STAT3 in CSC maintenance in PCa is intrinsically linked to its role as a critical component of IL-6 signaling. The importance of the IL-6/STAT3 axis has been linked to supporting CSC populations in a variety of cancers, including hepatocellular , breast in vivo xenograft growth better than AR targeting alone. Interestingly this mechanism occurs in cells with siRNA inhibition of the AR, further underscoring an inverse relationship between AR activity and the CSC/EMT phenotype [Interestingly, this requirement for IL-6 signaling, in PCa CSCs may underlie the observation that there is significant overlap or fluctuation between a CSC and EMT-like phenotype in may PCa cell lines. Indeed, many reports in PCa as well as other cancers have shown a correlation between expression of EMT and CSC markers within the same cells. For example, after androgen deprivation, both EMT and CSC populations increase in mouse prostates and PCa cells  and PCa henotype . In addihenotype . TGF-β1 henotype . Despite the numerous cell intrinsic pathways that endow tumor cells with their remarkable propensity for unrestricted growth, survival and dissemination, the interaction of cancer with their host and the microenvironment tumors create for themselves play equally important roles in the progression of disease. This is of course true for PCa, and newly emerging roles of the stromal cells, immune cells and secreted factors that mediate the interactions between these cell types and the tumor in the pre-metastatic and metastatic niches are being uncovered at a rapid rate.Salmonella vector inhibited PCa xenograft growth in vivo [Tumor mediated angiogenesis is a hallmark of solid tumors ; they re in vivo . Interes in vivo .in vitro and in vivo [As mentioned above, STAT3 also activates transcription of HIF-1α, which in itself is a transcriptional activator of VEGF, and STAT3 and HIF-1α cooperatively bind to the VEGF promoter to induce VEGF transcription . Inhibit in vivo ,90,91,92 in vivo . Interes in vivo .in vivo [in vitro and in vivo [Importantly, STAT3 signaling is not confined to regulating tumor cell intrinsic pathways that control the microenvironment; it is also a key player in the development of “reactive” or inflammatory stromal cells that have been shown to promote PCa tumor aggressiveness ,96. For in vivo  and loss in vivo . In addi in vivo . The act in vivo  and ocul in vivo  that ind in vivo . Moreove in vivo . Since SWhile primary prostate tumors are supported through interactions with fibroblastic stromal cells, tumor cells that metastasize modulate hematopoietic stroma in the bone to establish themselves within that niche. Over 80% of men that die from CRPC have tumors that have metastasized to the bone. These bone lesions are primarily osteoblastic and a variety of bone and cancer derived growth factors and chemokines play important roles in tumor establishment and bone destruction in this metastatic niche . One sigin vivo [in vitro and suppresses xenograft growth in vivo [An essential component of the tumor microenvironment that dictates cancer progression is the immune system. Indeed, evading detection and clearance by innate and adaptive immune cells is a hallmark of cancer  and STATin vivo . Like MDin vivo ,121. CCL in vivo ,122. The in vivo . Importain vivo models were not necessarily associated with inhibition of STAT3 itself. While many of these agents have anti-proliferative effects on PCa in vitro or in in vivo xenograft models in immunocompromised animals, few have gone on to show efficacy as single agents in patients. For example, the IL-6/JAK/STAT3 inhibitors siltuximab (CNTO 328) and ruxolitinib (INCB-018242) failed Phase II clinical trials due to a complete lack of PSA50 response [etc.) and cytokines in the IL-6 family. Importantly, in the studies aforementioned, most inhibition of PCa growth, migration, invasiveness or metastasis in in vitro or in vivo models was achieved using STAT3 specific inhibitors rather than inhibition of IL-6. However, combination therapy using IL-6 targeting with chemotherapy may prove more effective, as a recent phase I trial with siltuximab and docetaxel in combination showed robust PSA decline in the patients [Considering the wide array of signaling pathways that promote progression of CRPC that require STAT3, targeting this molecule for therapeutic benefit has been an intense area of investigation. This is true not only for PCa, but many other cancers as well. In response ,125. Thepatients . While tpatients , this mapatients . By contrast however, targeting Hsp27 has shown promise in PCa clinical trials in combination with prednisone in patients with metastatic CRPC; up to 71% of patients were progression free at 12 weeks and 50% showed 50% reduction in PSA when treated with the Hsp27 antisense OGX-427 and prednisone . While tA wide body of work spanning multiple solid tumor types, including PCa, have shown an indespensible and multifaceted role for STAT3 signaling in promoting tumor progression. Importantly, STAT3 has pivotal roles in re-activation of the AR, an essential step in the development of mCRPC. This can occur via STAT3 cooperation with other highly relevant oncogenic signaling cascades such as PTEN/PI3K/AKT and the integration of signals by STAT3 received from multiple growth factors, Src kinases and serine/threonine kinases that themselves have known roles in the progression of PCa. The diversity of signaling pathways in PCa tumor cells in which STAT3 acts as a major signaling node translates into it being a driving force in not only tumor proliferation and survival, but also in determining cell phenotype, behavior and interaction with stromal cells and the immune system. It is not surprising therefore that therapeutic targeting of STAT3 is under intense investigation for the treatment of mCRPC. However, due to the promiscuous nature of STAT3 signaling in so many cell types, the future success of potential therapies may possibly entail cell-specific delivery of STAT3 targeting or co-targeting with other known drivers of PCa progression to obtain optimal results. This concept has been explored to enhance immune responses in hematopoietic cancers, whereby targeting STAT3 specifically in TLR9 expressing cancer cells can be achieved using CpG based delivery of silencing oligos results in growth inhibition of myeloma and myeloid leukemia ,218. Ind


DNA damage is induced in many types of cells by internal and external cell stress. When DNA is damaged, DNA Damage Response (DDR) programs are activated to repair the DNA lesions in order to preserve genomic integrity and suppress subsequent malignant transformation. Among these programs is cell cycle checkpoint that ensures cell cycle arrest and subsequent repair of the damaged DNA, apoptosis and senescence in various phases of the cell cycle. Moreover, recent studies have established the cell differentiation checkpoint, the other type of the checkpoint that is specifically activated in the course of differentiation. We will discuss the evidences that support the link between DNA damage proteins and C2C12 cell differentiation. Genomic integrity is primordial to any organisms. It has been well illustrated that diverse stress from both intrinsic (ex. Reactive Oxygen Species (ROS)) and extrinsic (ex. ionizing radiation (IR), UV light, chemical) environment cause DNA lesions . When DNDifferentiation is the process in which cells become specialized from the precursor cells to specific cell type, such as neurons, lymphocytes and muscle through differentiation. A global reprogramming of gene expression and withdrawal from the cell cycle are required for the differentiation process . AlthougC2C12 cells have been widely used as an in vitro model to study myogenic differentiation process. These cells are derived from the mouse skeletal muscle C2 cell line, and they have similar characteristics to those of isolated human skeletal muscle cells ,11. MyogIt has been shown that during differentiation DNA Double Strand Breaks (DSBs) occur. For example, development of B lymphocytes requires the induction and consequent repair of DSBs during rearrangement of the antigen receptor genes . InteresNot only c-Abl, but also several DDR proteins have been implicated to be involved in the differentiation checkpoint, including ATM –25 and NLarsen et al. demonstrated that this DDR pathway is activated at the early stages of differentiation of C2C12 cells . PhosphoIn summary, myogenic differentiation consists of multistep, including appearance and repair of DSBs. When DSBs are generated, DDR proteins are activated to properly ensure the DNA repair before proceeding differentiation, guarantying the correct formation of differentiated cells without compromised genome. Of note, our recent findings showed that ATM inactivation. causes insufficient generation of dendritic cells from bone . Taken t


Cell polarity is a fundamental property of epithelial cells that confers spatial organization to signalling pathways regulating many aspects of cell physiology including survival, proliferation, and motility. Loss of epithelial organization and apical-basal polarity correlates with the acquisition of a malignant phenotype, and accumulating evidence suggests that loss of polarity signalling contributes to cancer progression. PAR3 is a multi-domain scaffold protein that functions as a central regulator of cell polarity, and is disrupted in multiple cancers. We recently reported that loss of PAR3 promotes tumor growth and metastasis in breast cancer. , 3 To prThis dual effect due to the loss of PAR3 is in some regards similar to a process termed compensatory proliferation, in which proliferation is induced to replace cells eliminated by apoptosis.  However,In addition to stimulating proliferation and tumor growth, loss of PAR3 promotes invasion and metastasis.  InterestIn summary, PAR3 acts as a tumor suppressor for breast cancer. Its function as a polarity scaffold spatially restricts multiple signalling pathways. Loss of this spatial organization deregulates proliferation and invasive programs to promote cancer progression when default apoptotic programs are inhibited. Figure .


Drug addiction is a term applied across substance use disorders (SUDs) and defined as a chronic, relapsing complex brain disease characterized by compulsive drug seeking, craving, loss of self-control, and impaired decision making  for nicotine  for alcohol and opioids   Enoch  and for α5-α3-β4 nicotinic cholinergic receptor subunit gene cluster on chromosome 15q, which sparked the discovery of several loci related to smoking and its more dire health consequences, such as lung diseases and peripheral artery disease  in addition to the more common α4/β2 nicotinic receptors, which are thought to mediate nicotine reward and dependence  and the interpeduncular nucleus (IPN), brain regions that express high levels of the Changeux , and whiDRD4 gene with individuals showing a greater risk for attention deficit hyperactivity disorder (ADHD) and for SUD. These individuals also show an increased sensitivity to both adverse as well as positive environmental factors resulting in either worse or better outcomes than those without the 7-repeat DRD4 VNTR  for SUD is mediated though their influence on brain development, connectivity, and function, which in turn influences brain responses. This more integrated perspective on the role of genes and gene regulation in SUD provides a better understanding of why there is significant comorbidity between SUD and mental illness, including not only the frequent co-occurrence of both of these disorders but also overlap in brain circuits, in risk alleles and in environmental insults. Particularly prominent are circuits involved with “self regulation and control,” which implicate frontal networks and those involved with saliency circuits, which implicate striatal and limbic regions . CNR1 is densely localized in the hippocampus, the amygdala, and the prefrontal cortex  and obesity risk , orexin, and corticotropin-releasing factor (CRF) systems and they can remain engaged even when the stressor is no longer present Koob . The CRFIt has become increasingly evident that epigenetic factors are instrumental in their ability to affect several genes simultaneously with or without the underlying genetic variation. Epigenomic regulation mediates cellular responses to external stimuli such as environmental toxins, infection, stress, and drugs of abuse. Epigenomic regulation of cocaine responses in brain reward circuitry has been particularly well studied Nester . ExposurIn summary, genetics and epigenetics have expanded our understanding of the neurobiological basis of SUDs and related disorders through an interdisciplinary process of human and molecular genetic research Fig. . The rem


Takotsubo cardiomyopathy (TCM) is a stress-induced cardiomyopathy that occurs primarily in postmenopausal women. It mimics clinical picture of acute coronary syndrome with nonobstructive coronary arteries and a characteristic transient left (or bi-) ventricular apical ballooning at angiography. The exact pathogenesis of TCM is not well recognized. Hereby we present an unusual case of TCM that presents with signs and symptoms of acute pericarditis and was also found to have a coexisting coronary muscle bridge on coronary angiography. We discuss the impact of these associations in better understanding of the pathogenesis of TCM. Takotsubo cardiomyopathy (TCM) or transient left ventricular apical ballooning syndrome is a novel type of cardiomyopathy that was first described in 1990 as a differential diagnosis for acute coronary syndrome in the absence of obstructive coronary artery disease , 3. AlthHereby we present an unusual case of TCM that presents with signs and symptoms of acute pericarditis and was found to have a coexisting coronary muscle bridge on coronary angiography (CAG), associations that might have an impact on better understanding of the pathogenesis of TCM.A 42-year-old Iranian woman was brought to the emergency department with a sharp, persistent pleuritic chest pain beginning after an emotional stress. She had neither remarkable past medical history for coronary risk factors nor any relevant past interventions. Physical examination revealed a heart rate of 85 beats per minute, blood pressure of 120/80 mmHg, and normal S1 and S2 with S3 gallop as well as a triphasic friction rub (one systolic and two diastolic sounds) at cardiac auscultation. An initial electrocardiogram (ECG) demonstrated 1-2 mm ST elevation in leads I, II, III, aVF, and V4–V6, as well as ST depression and PR elevation at aVR . A transAfter finding wall motion abnormality on TTE and because of persistent chest pain, despite the initial clinical diagnosis of acute pericarditis, the patient was immediately transferred to the cardiac catheter laboratory in order to look for any coronary artery disease requiring intervention. CAG demonstrated a severe muscle bridge at mid portion of the left anterior descending artery (LAD) and otherwise normal coronary arteries ; left veThe patient was treated with an angiotensin-converting enzyme inhibitor (captopril 6.25 mg three times per day), a diuretic (furosemide 20 mg three times per day), and a beta-blocker (carvedilol 3.125 two times per day). She also received aspirin for 4 days at high enough doses for treating pericarditis (650 mg three times per day). On serial ECG mild T-wave inversion was found in leads I, II, III, aVL, aVF, and V4–V6 at 2nd day of admission which then became deeper at 3rd day (Figures The patient's general condition considerably improved over the next few days and her chest pain was completely resolved. One week later, a subsequent TTE at discharge day revealed an improved left ventricular ejection fraction of 50–55% with no apical ballooning, no significant mitral regurgitation, and no pericardial effusion.In the present case, the fairly sudden onset of a sharp and pleuritic chest pain, the characteristic three-phasic pericardial friction rub, the mild pericardial effusion on TTE, and the significant clinical response to high-dose aspirin all are in favor of acute pericarditis. On the other hand, such features as the clear history of an emotional stress before the onset of chest pain, diffuse ST elevation on ECG with the patent coronary arteries at CAG, and the characteristic apical ballooning at TTE with spontaneous recovery are suggesting features of TCM. Finally, detection of coronary muscle bridge at CAG adds a third diagnostic issue to this patient's presentation.After the first formal description of TCM in 1990 in Japan , cases oAccording to the proposed diagnostic criteria of Mayo Clinic, absence of myocarditis is needed to confirm the diagnosis of TCM . MoreoveAnother interesting point in this case was coexistence of myocardial bridge, a congenital coronary anomaly characterized by systolic compression of a part of an epicardial vessel by a segment of overlying myocardium, and TCM. Association of myocardial bridge and TCM has been reported in some case reports and case series –30, whicIn conclusion, to the best of our knowledge, this is the first report of coexistence of myocardial bridging, TCM, and acute pericarditis in a single patient, which may suggest a potential role of both myocardial bridging and acute pericarditis in the pathogenesis of TCM.


GTPases and kinases are two predominant signaling modules that regulate cell fate. Dysregulation of Ras, a GTPase, and the three eponymous kinases that form key nodes of the associated phosphatidylinositol 4,5-bisphosphate 3-kinase (PI3K)/AKT/mTOR pathway have been implicated in many cancers, including pancreatic cancer, a disease noted for its current lack of effective therapeutics. The K-Ras isoform of Ras is mutated in over 90% of pancreatic ductal adenocarcinomas (PDAC) and there is growing evidence linking aberrant PI3K/AKT/mTOR pathway activity to PDAC. Although these observations suggest that targeting one of these nodes might lead to more effective treatment options for patients with pancreatic and other cancers, the complex regulatory mechanisms and the number of sequence-conserved isoforms of these proteins have been viewed as significant barriers in drug development. Emerging insights into the allosteric regulatory mechanisms of these proteins suggest novel opportunities for development of selective allosteric inhibitors with fragment-based drug discovery (FBDD) helping make significant inroads. The fact that allosteric inhibitors of Ras and AKT are currently in pre-clinical development lends support to this approach. In this article, we will focus on the recent advances and merits of developing allosteric drugs targeting these two inter-related signaling pathways. Dysregulation of cellular signaling pathways that involve phosphoryl transfer through GTPase and kinase activities is a key step in the initiation and development of neoplasms  form a key node in the PI3K/AKT/mTOR pathway, regulating numerous biological processes through phosphoryl transfer  co-structure  are known to cause a variety of cancers, with K-Ras being the most frequently mutated isoform in pancreatic cancer . As mutations in Ras are common to a variety of cancers many different strategies that aim to down regulate constitutively active mutant forms have been explored  . This observation prompted Ostrem and co-workers to use mass spectrometry to screen a library of fragment-like compounds that contain reactive sulfur moieties for ones which covalently modified K-RasTable 1(A) Allosteric inhibitors currently either FDA approved or in clinical trials that target major nodes of the PI3K/AKT/mTOR pathway. (B) A selection of compounds reported as allosteric inhibitors of Ras and major nodes of the PI3K/AKT/mTOR pathway.The PI3K/AKT/mTOR pathway forms one of many mechanisms that regulate progression though the cell cycle and cellular apoptosis  all containing an N-terminal PH domain that binds PIP3, a central AGC kinase domain, and a hydrophobic C-terminal domain. After recruitment to the plasma membrane through binding PIP3, AKT is activated by two phosphorylation events: one on its activation loop  by membrane-associated PDK1 and the other on the C-terminal hydrophobic motif by mTORC2 (Ser473)  is a member of the serine/threonine phosphatidyl inositol 3′ kinase-related kinase family (PIKK) that uniquely phosphorylates protein substrates and is sensitive to rapamycin, a large macrocyclic compound that is synthesized by Being a member of the PIKK family, it is unsurprising that many compounds developed to target the ATP-binding site of PI3K can also inhibit mTOR  and the complex nature of its multi-protein assembly, there is likely significant opportunity to develop selective drugs that modulate mTORC1/mTORC2 activities and hence deliver more fine-tuned treatments for cancer.Allosteric compounds are making it into mainstream drug development due in part to recent advances in structural biology and biophysical techniques, FBDD, and computational chemistry. As summarized in this mini-review, recent drug discovery programs have shown that allosteric inhibitors may have an edge over more common active site-competitive inhibitors, particularly when targeting specific members of a family of related proteins. An added benefit to this approach appears to be the reduction in likelihood of obtaining unexpected and undesired results like that seen in hyperphosphorylated AKT. The emerging knowledge of allosteric regulatory mechanisms of therapeutically important targets, such as PI3K, may aid in future allosteric drug development, although caution should be noted, as some targets are likely to remain intractable to allosteric inhibition by their intrinsic nature.Unlike the discovery and development of drugs that target active sites, allosteric drug discovery may present more of a challenge as methodologies for detection and analysis of hit and lead compounds are still maturing. However, emerging technologies, such as micro-scale thermophoresis for compound screening and diversity oriented synthesis for library design, may help accelerate the discovery and development of drugs that allow effective treatment of currently incurable cancers such as PDAC.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Autophagy is an intracellular pathway for bulk protein degradation and the removal of damaged organelles by lysosomes. Autophagy was previously thought to be unselective; however, studies have increasingly confirmed that autophagy-mediated protein degradation is highly regulated. Abnormal autophagic protein degradation has been associated with multiple human diseases such as cancer, neurological disability and cardiovascular disease; therefore, further elucidation of protein degradation by autophagy may be beneficial for protein-based clinical therapies. Macroautophagy and chaperone-mediated autophagy (CMA) can both participate in selective protein degradation in mammalian cells, but the process is quite different in each case. Here, we summarize the various types of macroautophagy and CMA involved in determining protein degradation. For this summary, we divide the autophagic protein degradation pathways into four categories: the post-translational modification dependent and independent CMA pathways and the ubiquitin dependent and independent macroautophagy pathways, and describe how some non-canonical pathways and modifications such as phosphorylation, acetylation and arginylation can influence protein degradation by the autophagy lysosome system (ALS). Finally, we comment on why autophagy can serve as either diagnostics or therapeutic targets in different human diseases. Autophagy is an evolutionarily conserved eukaryotic process that can be initiated in response to both external and intracellular factors, including amino acid starvation , growth Unfolded or misfolded cellular proteins are generally degraded by the proteasome; however, proteasome activity can be inhibited by large protein polymers due to their inability to pass through the narrow proteasome channel; thus, autophagy is stimulated to eliminate those protein complexes , 15. MacAutophagy occupies a central position in the maintenance of cellular homeostasis by directing protein degradation, and the process adapts cells to adverse micro-environmental conditions. Accordingly, unraveling more details about autophagic protein degradation pathways may not only improve our understanding of cell metabolism and fate, but may also help us understand a diverse range of human diseases. In this review, we focus on diverse autophagy-mediated protein degradation processes and their regulation.The ubiquitin proteasome system (UPS) and the autophagy-lysosome system (ALS) are alternative ways to categorize protein degradation based on the functions of the proteins being degraded. It is well known that large protein complexes cannot fit into the narrow proteasomal channels and are thus directed to lysosomal disruption , 15.There are three types of proteins in cells that are degraded by autophagy Figure ; the firChaperone-mediated autophagy (CMA) is a type of autophagy that degrades soluble or unfoldable proteins in a molecule-by-molecule fashion Figure . In contHSC70 is a member of the heat shock protein 70 family and is located in the cytosol or the lumen of lysosomes . ProteinThe criterion for the protein to be a putative CMA substrate is the presence of a peptide sequence that is biochemically related to the KFERQ motif. In brief, the motif is flanked by a glutamine (Q) and contains one acidic residue (D or E), a basic residue (K or R), a hydrophobic residue , and a fifth residue that can be basic or hydrophobic but cannot be negatively charged . In partRecent studies have indicated that oxidative stress can activate the CMA pathway and that this potentiation occurs at least partially because oxidized proteins are more easily unfolded to facilitate translocation into the lysosomal lumen. In addition, lysosomes are more active under oxidative stress. For example, the expression of LAMP2A and Lys-HSC70 increased in the lysosomal membrane and lumen during oxidative stress . MoreoveIn addition to oxidative stress and oxidation, other forms of modifications such as phosphorylation, ubiquitination and acetylation can participate in the regulation of CMA. When Phosphoprotein enriched in diabetes (PED) was phosphorylated at Ser104 or Ser116 near the KFERQ-like motifs, its interaction with HSC70 was reduced, thereby precluding its degradation . In contAlthough the precise mechanism by which the modifications affect the interaction between the protein and HSC70 is not yet clear, many CMA substrates were identified during the last decade and were listed in J. Fred Dice's article . In thisAutophagy receptors and the ATG8 family are two primary elements in selective macroautophagic protein degradation. Autophagy receptors bind cargo and the ATG8 family plays a pivotal role in the selective macroautophagy process by promoting the entry of cargo receptors into the autophagy cascade via interaction with the LC3-interacting regions (LIR) of the autophagy receptor , 59. TheTo date, several autophagy receptors that mediate protein degradation have been identified, including TOLLIP, SQSTM1/p62, NDP52, OPTN and NBR1. These receptors contain four different ubiquitin-binding domains: the coupling of ubiquitin conjugation to endoplasmic reticulum-associated degradation domain (CUE), the ubiquitin-associated domain (UBA), the ubiquitin-binding zinc finger domain (UBZ) and the ubiquitin binding in ABIN and NEMO domain (UBAN), along with one or two LIR domains Figure  63, 64], 6463, 6Selective protein degradation by macroautophagy has gained increasing attention due to increased studies of the roles of SQSTM1 in ubiquitinated protein degradation . Recent Similar to UPS, there is strong evidence supporting the recognition of ubiquitinated proteins by receptors, which helps to include them in the macroautophagy cascade Figure  63]. Pr. Pr63]. There are two E1 activating enzymes, 40 E2 conjugating enzymes and more than 500 E3 ligases for ubiquitination in mammals . DistincMoreover, the post-translational modifications of autophagy receptors can influence their interaction with ubiquitinated substrates. The phosphorylation of p62 S403 in its UBA domain by casein kinase 2 (CK2) enhances its binding affinity to ubiquitinated proteins and then promotes the autophagic clearance of the substrate . HoweverUnlike the ubiquitination in the ALS, deubiquitinating enzymes that negatively regulate autophagic protein degradation in mammals have rarely been reported. To date, only one deubiquitinating enzyme, USP36, has been confirmed. USP36 can inhibit selective macroautophagy by preventing the accumulation of ubiquitinated proteins, but the specific substrates of USP36 (whose targeting to autophagosomes is affected) are not yet known . TherefoSubstantial evidence has implicated ubiquitin as the culprit responsible for macroautophagic protein degradation. However, recent results argue against the possibility that ubiquitin is the ubiquitous code in selective macroautophagy Table . For exaIn addition to the direct interaction with an autophagy receptor or ATG8 family member Figure , proteinMacroautophagy and CMA are two distinct mechanisms of protein degradation; however, crosstalk between these pathways has been reported. BAG3 coupled with HSC70, which participates in CMA by recognizing the KFERQ motif in substrates, can release HSC70 substrates to the dynein motor complex, thereby mediating aggresome targeting and the macroautophagic degradation of chaperoned substrates . This prIn addition to the crosstalk between different autophagy pathways, autophagy and proteasome pathways have been demonstrated to work complementarily to degrade proteins under some circumstances. For example, when the proteasome system cannot degrade large protein aggregates, HSP90 and Poh1 help the aggregates dissociate from 20S proteasomes while macroautophagy is activated to facilitate clearance of these protein aggregates . TherefoMoreover, the proteasome and lysosome are believed to degrade misfolded proteins with the help of heat shock proteins, and it was therefore believed that these pathways are both activated to eliminate misfolded proteins that are able to pass through the narrow proteasome channel. Interestingly, the SOD1 G93A mutant was reported to be ubiquitinated by E3 ligase gp78, which targets the protein for proteasome degradation . HainingPrevalent human diseases, including neurodegenerative diseases and carcinomas, have been reported to be attributable to dysfunction of protein degradation; therefore, certain functional elements in ALS or UPS tend to be used as either diagnostic or therapeutic targets in the treatment of these diseases.A recent finding has indicated that lysosomal proteins LAMP1 and LAMP2 and the autophagy protein LC3B in human cerebrospinal fluid (CSF) may be potential novel biomarkers for Alzheimer disease ; howeverin vitro and in vivo models of myeloma [Similar to ALS, abnormal UPS, including its proteasomes and proteases, is an indicator and target of numerous human diseases . The ant myeloma .Proteasome inhibitors such as Bortezomib and DUB inhibitor PR-619 can trigger autophagy, although the mechanisms of these activities are not yet fully understood , 183. InAutophagic degradation has been demonstrated to be selectively and precisely regulated. However, details regarding this type of degradation pathway remain largely unknown. Ubiquitination has been shown to accelerate autophagic protein degradation by interacting with the ubiquitin-binding domains in autophagy receptors. Interestingly, recent studies showed that, compared with non-arginylated BiP and non-acetylated Huntingtin proteins, arginylated and acetylated proteins strongly bind p62 to undergo degradation by autophagy, though the precise mechanisms remain unknown , 93. ThuLong noncoding RNAs (lncRNAs), defined as transcripts longer than 200 nucleotides, are a currently popular study topic in research due to their far-ranging functions in regulating protein-protein and protein-RNA interactions , 188. ThEctopic accumulation and distribution of functional proteins are adverse to human health. Although autophagic removal of insoluble toxic proteins or oncogenic proteins could relieve neurological or tumorous symptoms respectively, specific and effective targeting by autophagy drugs is insufficient compared with numerous UPS intervening agents. Therefore, the questions of whether any drugs that can selectively modulate protein and autophagy receptor interactions or whether E3 ligase inhibitors or DUB inhibitors can control the autophagic degradation of certain protein targets remain to be further determined. Further investigation of autophagic protein degradation mechanisms may help elucidate these questions and provide more unique therapeutic strategies to treat human diseases.


Internalization and degradation of live Bb within phagosomal compartments of monocytes, macrophages and dendritic cells (DCs), allows for the release of lipoproteins, nucleic acids and other microbial products, triggering a broad and robust inflammatory response. Toll-like receptors (TLRs) are key players in the recognition of spirochetal ligands from whole viable organisms . Herein we will review the role of endosomal TLRs in the response to the Lyme disease spirochete. Borrelia burgdorferi (Bb), the causative agent of Lyme disease  2/TLR1 heterodimers on the surface of phagocytic cells to induce the production of inflammatory cytokines , including spirochetal lipoproteins and nucleic acids. The interaction between spirochetal PAMPs and the TLRs triggers a variety of intracellular signaling pathways leading to the production of various cytokines, including type I interferons   was demonstrated to directly bind Bb and the presence of complement enhanced spirochetal binding  (integrin αScavenger receptors comprise a group of unrelated transmembrane surface molecules with relatively promiscuous ligand binding such as scavenger receptor A (SR-A), MARCO (Macrophage receptor with Collagenouse structure) and CD36  mice infected with Bb , psoriasis, rheumatoid arthritis, Sjögren's syndrome and others (Demaria et al., erythema migrans and Lyme carditis often present within the first few weeks of intection, Lyme arthritis more often presents several weeks after the initial infection in untreated patients and can persist even after antibiotic treatment (Kean and Irvine, One of the more puzzling aspects of Lyme disease is the persistence, in some patients, of musculoskeletal symptoms following Bb infection, and their refractoriness to rapid improvement despite proper antibiotic treatment (Bockenstedt et al., Naked pathogen-derived nucleic acids present in the extracellular space upon release from damaged or disintegrated microbes or the infected host cells, may be ultimately degraded by extracellular DNases and RNases before they can access the endolysosomal compartment of other immune cells (Brencicova and Diebold, TLR9 is another endosomal TLR that has been linked to autoimmunity (Theofilopoulos et al., While TLRs are capable of sensing pathogenic materials, defective TLR signaling can hinder activation of the adaptive response. On the other hand, excessive response of TLRs and production of cytokines may increase the disease state (Kean and Irvine, erythema migrans, systemic inflammation, Lyme arthritis, Lyme carditis, and neuroborreliosis (Radolf et al., Bb produces many symptoms within the human host, including Borrelia infection can also lead to Lyme carditis, a dangerous condition that can in rare instances can lead to sudden death CDC, . The mecTLR2 gene, specifically Arg753Gln, are less responsive to PAMPs derived from Bb (Schwartz and Cook, With recent progress in DNA analysis, the extremely polymorphic genes of TLRs can finally be understood. The genetic variability in TLRs can result in functional deficiency, which ultimately leads to immunodeficiency syndromes. The TLR1 1602S polymorphism, found predominantly in European-Caucasian populations, has been correlated with low expression of TLR1 at the surface membrane (Kean and Irvine, TLR7 have been associated with more severe Hepatits C viral infection (Lin et al., TLR8 gene have been linked to increased susceptibility to bacterial infections (Davila et al., TLR signaling alterations have been linked to more severe clinical manifestations in response to bacterial, fungal and viral infections (Frazao et al., It is important to note that despite the large number of TLR gene mutations found in the general population, most affected individuals will not suffer life threatening complications or even more severe infections than their unaffected counterparts (Netea et al., Future studies will attempt to elucidate the relationship between TLR mutations and the short and long-term outcome of human Lyme disease, particularly as to changes in these key innate immune receptors have a role in patients with prolonged, antibiotic refractory Lyme disease.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Gliomas are a highly heterogeneous group of brain tumours that are refractory to treatment, highly invasive and pro-angiogenic. Glioblastoma patients have an average survival time of less than 15 months. Understanding the molecular basis of different grades of glioma, from well differentiated, low-grade tumours to high-grade tumours, is a key step in defining new therapeutic targets. Here we use a data-driven approach to learn the structure of gene regulatory networks from observational data and use the resulting models to formulate hypothesis on the molecular determinants of glioma stage. Remarkably, integration of available knowledge with functional genomics datasets representing clinical and pre-clinical studies reveals important properties within the regulatory circuits controlling low and high-grade glioma. Our analyses first show that low and high-grade gliomas are characterised by a switch in activity of two subsets of Rho GTPases. The first one is involved in maintaining normal glial cell function, while the second is linked to the establishment of multiple hallmarks of cancer. Next, the development and application of a novel data integration methodology reveals novel functions of RND3 in controlling glioma cell migration, invasion, proliferation, angiogenesis and clinical outcome. de novo, it is common that low-grade gliomas progress to the more aggressive form known as glioblastoma. In this article, we have shown that by combining advanced network biology approaches with the right experimental models, we are able to reveal novel regulatory circuits controlling multiple hallmarks of glioma. Through analysis of multiple network models representing protein-protein interaction or gene co-expression data we have revealed a switch in the role of regulatory Rho GTPases between low and high-grade gliomas. Amongst these, we show that RND3 is up-regulated in glioblastomas and is a key regulator of tumour proliferation, migration and invasion. We confirm that expression and genomic copy number of RND3 are predictive of clinical outcome, suggesting that changes in the activity of this particular Rho GTPase could be an early event associated to transformation and tumour expansion.Gliomas are aggressive brain tumours that are invasive, heterogeneous, refractory to treatment and show poor survival rates. Surgical resection and chemotherapy can increase patient survival but ultimately the disease is fatal. Multiple grades of glioma exist, with lower grades associated to better prognosis. While the majority of high-grade gliomas occur  De novo GBM constitute the majority of grade IV glioma and are powerful inducers of angiogenesis, highly proliferative and invasive. They are largely resistant to treatment and have poor prognosis with two years survival rates as low as 3.3% [Gliomas are brain tumours originating from the glial cells and neural stem cells that surround and support neurons . They ar as 3.3% .A number of studies have identified key genomic alterations in GBM able to induce transformation in non-tumorigenic cells such as mutations within EGFR  6] and  and 6] ain vivo and in vitro glioma models.The complexity of the factors involved in the biology of gliomas makes it difficult to develop a comprehensive model underlying GBM progression. Here we address this important challenge by integrating functional genomics datasets representing existing knowledge, clinical studies and We first show that network modules derived from a comprehensive integration of protein interaction databases and defined by a high density of genes differentially expressed between low and high-grade gliomas are consistent with the hypothesis that Rho GTPases may be part of a key regulatory mechanism controlling hallmarks of high-grade glioma. A key feature of GBM is invasion of tumour cells into the surrounding brain tissue and members of the Rho GTPase family known to control actin cytoskeleton dynamics and cell migration have been implicated in the survival and invasion of tumour cells  13], , Further characterisation of a high-grade glioma transcriptional network highlights a pivotal role of the Rho GTPase RND3  in controlling tumour proliferation, migration and invasion. Ultimately, the clinical relevance of this regulatory network is proving that copy number variation in the RND3 gene is predictive of clinical outcome.Our study is based on a complex data analysis workflow which includes several complementary reverse engineering techniques to address the important challenge of generating and validating hypotheses outlining the main factors underlying the control and maintenance of glioma stage. The strategy we followed, which is summarised in The workflow consisted of five distinct but interconnected steps.Step 1- Integration of protein-protein interaction networks with gene expression data derived from a human clinical study: This represents low and high-grade gliomas and were used to modularise a large network of known human protein-protein interactions (PPi). The analysis of these modules identified a unique sub-network of regulatory factors, which represented a number of Rho GTPases.Step 2- Revealing the linkage between subsets of Rho GTPases and glioma grade: Genome scale gene expression data were used to construct co-expression networks centred on the regulatory Rho GTPases identified in step 1. This revealed a switch in activity of Rho GTPases between grade II and grade IV glioma.in vivo model:Step 3- Reverse engineering of regulatory networks from an We used the experimental glioma model in the chick chorioallantoic membrane  which rein vitro and in vivo).Step 4- Validation step: The predictions from the network analysis were validated through functional assays  mainly comprised genes up regulated in grade IV gliomas and were enriched in typical effector functions in cancer. More precisely, the largest of the effector modules (M2) was enriched in genes linked to apoptosis, blood vessel development and inflammatory response and included the oncogenes JUN, FOS and BCL3 (for reviews of known oncogenes see ). ). M2-4)n glioma  13] [19 [19M2-4n glioma  15]  and somatic mutations [SNP:ID] or (E) copy number variation, gene expression [mRNA] and somatic mutations from the TCGA database to RND3 expression. Percentage of variance in RND3 expression explained: D– 13.97%, E– 21.04%. Plus or minus symbols indicate sign of the Spearman correlation value between RND3 and CNV/mRNA expression.(TIF)Click here for additional data file.S1 TextThis document describes the background, methodology and performance of the novel multi-level data integration approach used in this study to generate the networks in (DOCX)Click here for additional data file.S1 TableSignificant enrichment indicated by bold text .(TIF)Click here for additional data file.


Hepatocellular carcinoma (HCC) is the second most common cause of cancer-related death worldwide , generalPhenotypic intratumor heterogeneity in HCC with respect to morphology and differentiation grades within the same tumor is a well-known phenomenon to surgical pathologists. So far, genetic heterogeneity and the heterogeneity of biomarker expression in the surgical HCC specimens have not been systematically analyzed. However, to improve clinico-pathological classification systems and for the stratification of targeted therapies, it seems crucial to comprehensively characterize intratumor heterogeneity.TP53 and CTNNB1). Combining Sanger sequencing with deep sequencing techniques facilitated the discovery of low frequency mutations and mitigated the effect of wild-type contamination. A mean of 7 regions per tumor was sequenced , and genetic intratumor heterogeneity was found in 22% of cases. Thus, already analysis of the two main HCC driver genes clearly revealed that mutations are not homogeneously present in all regions of an individual tumor. This was found especially for CTNNB1 mutations, but also for TP53 mutations. The thorough dissection of morphologic, immunohistochemical and genomic intratumor hetero-geneity in our study illustrates that the primary co-existence of different growth patterns can be associated with divers biomarker expression and TP53 or CTNNB1 gene mutations among wild type tumor cells. Somatic mutations of various other genes are described in liver cancer, e.g. AXIN1 , ARID and MLL genes (epigenetic modifiers), CDKN2A and IRF2 (cell cycle regulators interrelated with TP53) or TERT promoter mutations [In a systematic analysis of 23 treatment-naïve unifocal HCC, we investigated individual tumors for morphologic, immunohistochemical and genetic intratumor heterogeneity as well as the association of these three features [utations . AlthougIntratumor heterogeneity has major implications for diagnosis and therapy of many solid cancers, indicating that a single tumor biopsy might not provide sufficient informative value regarding the molecular characteristics of the whole tumor. Primary renal cell cancer, as has been shown by Gerlinger et al. , displayThe histopathologic and molecular classification of a tumor, often determined by the mutational status of a certain target, has therapeutic implications, e.g. in colorectal cancer (EGFR) or gastrointestinal stroma tumors (c-KIT). In the era of targeted therapies, intratumor heterogeneity is a major challenge to successful cancer therapy since it may result in primary resistance or early evasion of treatment to chemotherapeutic or molecular targeted substances. In melanoma, resistant tumor clones tend to evade chemotherapies and overgrow due to a forced selection process as well as an adaption process during tumor evolution. Therapy-induced inflammatory processes result not only in phenotypic plasticity of the tumor and its genomic landscape, but also alter the composition of tumor-associated immune cells. This recomposition of the microenviroment can contribute to cancer evolution and therapy resistance .In conclusion, determining the degree of intratumor heterogeneity might be seen as a biomarker by itself and have prognostic value for disease progression . With th


Saccharomyces cerevisiae) (SIRT1) and the cell survival mechanisms of autophagy that can be coupled to the activity of the mechanistic target of rapamycin (mTOR). Targeting miRNAs that oversee SIRT1 activity offers interesting prospects for the translation of these pathways into efficacious clinical treatment programs for a host of disorders. Yet, as work in this area progresses, a number of challenges unfold that impact whether manipulation of non-coding RNAs and SIRT1 can finely guide stem cell renewal and differentiation to reach successful clinical outcomes.Small non-coding ribonucleic acids (RNAs), known as microRNAs (miRNAs), are now becoming recognized as significant agents that can affect the onset and progression of numerous disorders throughout the body. In particular, miRNAs also may determine stem cell renewal and differentiation. Intimately tied to the ability of miRNAs to govern stem cell proliferation are the proliferative pathways of silent mating type information regulation 2 homolog 1 ( Stem cells are increasingly being considered for the development of novel strategies for multiple disorders throughout the body that can affect the nervous system, cardiovascular system, immune system, metabolism, and cancer. One of the challenges for applications that rely upon stem cell proliferation and differentiation is the protection and maintenance of stem cell populations. For example, specific pathways, such as the mechanistic target of rapamycin (mTOR), can be critical for stem cell proliferation . In the Saccharomyces cerevisiae) (SIRT1) activity that also is vital for stem cell proliferation [Interestingly, loss of mTOR activity can promote the induction of autophagy  and leadferation . In humaferation . SIRT1 aferation ,14. Duriferation  and incrferation . SIRT1 cferation , preventferation , and impferation . SIRT1 cferation . Differeferation . As a prferation , promoteferation , and fosferation .One strategy that may successfully regulate SIRT1 activity and stem cell proliferation for effective translation into clinical treatment programs may involve the modulation of microRNAs (miRNAs). MiRNAs are composed of 19-25 nucleotides and are small non-coding ribonucleic acids (RNAs). MiRNAs oversee gene expression by silencing targeted messenger RNAs (mRNAs) translated by specific genes. These small non-coding ribonucleic acids may play an important role to influence stem cell proliferation and cellular differentiation. For example, over-expression of miR-381 can lead to neural stem cell proliferation and prevent differentiation into astrocytes . In regaTargeting miRNAs provides an intriguing format for the control of stem cell proliferation and differentiation through pathways that involve SIRT1. Yet, several considerations must be addressed for the development of novel strategies with stem cells, miRNAs, and SIRT1. For example, the cellular level of activity of SIRT1 that is controlled by miRNAs may present an important caveat for the development of strategies for clinical disorders, since the presence of SIRT1 has the capability to either promote or retard stem cell proliferation and differentiation. To a similar degree, the level of SIRT1 activity can ultimately influence cellular survival. Sufficient SIRT1 activity is required for cellular cardiovascular protection -35 and n


Glutamatergic neurons of the mammalian cerebral cortex originate from radial glia (RG) progenitors in the ventricular zone (VZ). During corticogenesis, neuroblasts migrate toward the pial surface using two different migration modes. One is multipolar (MP) migration with random directional movement, and the other is locomotion, which is a unidirectional movement guided by the RG fiber. After reaching their final destination, the neurons finalize their migration by terminal translocation, which is followed by maturation via dendrite extension to initiate synaptogenesis and thereby complete neural circuit formation. This switching of migration modes during cortical development is unique in mammals, which suggests that the RG-guided locomotion mode may contribute to the evolution of the mammalian neocortical 6-layer structure. Many factors have been reported to be involved in the regulation of this radial neuronal migration process. In general, the radial migration can be largely divided into four steps; (1) maintenance and departure from the VZ of neural progenitor cells, (2) MP migration and transition to bipolar cells, (3) RG-guided locomotion, and (4) terminal translocation and dendrite maturation. Among these, many different gene mutations or knockdown effects have resulted in failure of the MP to bipolar transition (step 2), suggesting that it is a critical step, particularly in radial migration. Moreover, this transition occurs at the subplate layer. In this review, we summarize recent advances in our understanding of the molecular mechanisms underlying each of these steps. Finally, we discuss the evolutionary aspects of neuronal migration in corticogenesis. The mammalian neocortex is a highly organized structure underlying higher brain functions such as cognition, learning, and memory. It consists of a six-layer structure with an inside-out pattern, which is formed by radial migration of neuroblasts that continuously bypass the preceding differentiated and migrated neurons. Because neurons are born in the deeper part of the developing brain and migrate toward the pial surface, proper regulation is crucial, and impairment of this process results in various disorders such as brain malformation or psychiatric diseases. Our understanding of how this mammalian-specific complex structure is organized has advanced substantially in the last 20 years. In this review, we summarize the molecular pathways underlying how newly developed neurons travel from their birth to the terminus by dividing the process into four parts, as shown in Figure Neural progenitor cells (NPCs) of the glutamatergic neurons of the mammalian neocortex proliferate via symmetrical division of neuroepithelial cells in the early developmental stage. During development, neuroepithelial cells become radial glia (RG) cells by expressing marker proteins that are characteristic of astrocytes, including glial fibrillary acidic protein (GFAP), astrocyte-specific glutamate transporter (GLAST), the brain lipid binding protein (BLBP), and tenascin C (TNC) around the onset of neurogenesis. RG cells have a long basal process that extends to the pial surface, and they start producing neurons by asymmetrical division while maintaining symmetrical division , Ascl1, and Dll1, a key ligand for activating Notch signaling  cortex. Upon differentiation to IP cells, upregulated Ngn2 repressed Sox2 followed by cyclin D1 de-repression, and promote IP proliferation  transcription factor, an early born postmitotic neuronal marker  cell competency  and Gli3. Brn1/2 is a crucial regulator of the production of upper-layer neurons, and its expression in VZ progenitors is essential for the transition from early to mid-neurogenesis  complex 2(PCR2), exhibited the same phenotype of prolonged neurogenic phase of NPCs and delayed onset of the astrogenic phase, as depletion of Ring1B at the same stage (E12.5)  are an important component of cytoskeletons and are vital for the organization of the centrosome and mitotic spindle, which are also crucial for the maintenance of NPCs. EFHC1 is a protein containing a single EF-hand motif, a CaLGN (G protein regulator) gene or manipulation of the mouse Inscuteable(mInsc) gene expression level leads to a disruption of the balance between proliferation and differentiation of NPCs , also called elevator movement 18.5, which was caused by impairment of cell cycle exit and reduced proliferation of NPCs , sonic hedgehog (shh), Wnt, bone morphogenetic proteins (BMPs), colony stimulating factor-1 (CSF-1), and interleukin (IL) 34 are involved in the regulation of progenitor self-renewal and differentiation. Targeted disruption of the docking protein FRS2α, a major mediator of FGF signaling, leads to severe impairment of cerebral cortical development with thinner cerebral cortices than wild-type (WT) cells, reduced proliferation and differentiation of Tbr2- positive bIPs  influences the proliferation and differentiation of progenitor cells in the VZ  secretion and impairment of switching from symmetric to asymmetric division, thus leading to a decrease in neuron and IP production  at E12.5 contribute to maintain the neocortical progenitor pool  shape with multiple neurites and migrate in a MP migration mode in random directions , a proneural transcription factor responsible for glutamatergic neuronal differentiation from NPCs, has been found to also play an important role in the MP-BP transition via direct transcriptional activation of the small GTP-binding protein Rnd2, an atypical Rho-GTPase protein that inhibits RhoA activity and regulates actin cytoskeleton  family, is known as a putative transcription factor and silencing of this gene is associated with neurodevelopmental disorders and intellectual disability (ID)  expressed in the developing cerebral wall are also involved in the regulation of radial migration , also known as the Ras superfamily, comprise more than 150 small G proteins that can be divided into five subfamilies: Ras, Rho, Rab, Arf, and Ran   are GEFs for Arf1. ArfGEF2 null mice develop PH and exhibit neuronal migration defects of the developing cortex  is a human cortical malformation disease associated with mutations in the As described above, small G-proteins involved in the regulations of membrane trafficking, cytoskeletal organization and cell adhesion play critical roles in dynamic morphological changes during the MP-BP transition during radial migration of neuroblasts. It is undeniable that many causative genes of neurodevelopmental diseases belong to small G-protein family.Lissencephaly-1 (LIS1) is the first identified gene responsible for type I lissencephaly  is a gap junction protein that assembles a hemichannel of large- diameter channels in gap junctions. It has been reported that Cx43 is necessary for neuronal migration, especially the MP-BP transition independent of its channel-forming activity  is a type I transmembrane glycoprotein, and its proteolysis product Aβ accumulates in neurons in Alzheimer's disease. However, the cellular function of APP in normal neuronal development remains unknown. Young-Pearse et al.  found thRecently, it was reported that serotonin 6 receptor (5-HT6R), a G protein-coupled receptor (GPCR), is critical for the MP-BP transition and locomotion , which is characterized by macrocephaly, craniofacial dismorphism, hypotonica, cognitive disability, and intrac epilepsy  is another polarity kinase involved in the regulation of centrosome dynamics  is known to regulate axogenesis of dissociated cultured neurons  mice showed that RhoA activity is dispensable for migrating neurons, but is required for proper formation of the RG scaffold , as mentioned above, have distinct roles in each step although they belong to the same protein family. This suggests that a distinct set of genes participate in regulation of locomotion to fulfill the switching of migration mode during cortical evolution, as we discuss later.Finally, when migrating neurons arrive at their final destinations, they terminate migration and shift to terminal translocation and maturation by extending their axons and dendrites. This termination step should be executed by at least three distinct behaviors of locomoting neurons: locomotion termination, detachment from the RG fiber, and anchoring to the MZ for terminal translocation Figure .Lmx1a -GPR56 signaling for PBM integrity to prevent neuronal over-migration of preplate neurons. The same research group found that α3β1 integrin functions together with GPR56 to produce a proper stopping signal  lissencephaly with a defect in basement membrane (BM) integrity. This includes over-migration of the early born neuronal population as well as late-born locomoting neurons. Many factors have been reported to be involved in this over-migration defect, including the LIM homeobox gene SPARC-like 1 (SC1), a member of the SPARC family of ECM proteins, is involved in detaching locomoting neurons from the RG fibers, and is expressed at the top and bottom of the RG cell surface. SC1 was identified via antigen screening for radial glial immunoreactive monoclonal antibodies. SC1 possesses an anti-adhesive activity that may contribute to detaching the locomoting neurons from the RG fibers at their final step of locomotion. The absence of SC1 results in defective termination of locomotion and the final positioning of neurons, suggesting that anti-adhesive signaling at the termination phase is essential  layer, but are stacked just below the SP Figure . Howeverin vitro were identified for axon determination, more signaling pathways are required for leading process formation, suggesting the importance of in vivo signaling from the extracellular environment.Our review of recent progress in understanding the molecular pathways involved in radial neuronal migration of glutamatergic neurons during corticogenesis revealed that proper regulation of the various signaling pathways operating inside or outside of newborn neurons (neuroblasts) is critical for each step of neuronal migration. In general, accurate control of developmental timing for gene expression, translational control, and modification of protein activity is required for embryonic morphogenesis. To understand the regulatory mechanisms, it is important to determine how extrinsic signaling cues translate into intracellular signals. Newborn cortical neurons execute a dynamic morphological change from MP cells to BP cells. During this step, axons are first determined, followed by formation of thick leading processes, before making the transition to locomotion mode. Although the many factors involved in regulation of polarity formation in vivo neuronal activities and the effects of activity manipulation. To investigate protein-protein interactions, or protease activity on the ECM, the use of fluorescent probes such as the proximity ligation assay and protease imaging could be effective techniques.So far, explorations of gene function have been most commonly performed using knockdown or overexpression experiments, by delivering the DNA constructs with the IUE technique or via genetic modification of the gene locus to establish conventional and conditional knockout mice. However, to achieve more comprehensive understanding of these complex processes beyond determination of individual gene function, we could apply these conventional techniques in combination with more advanced techniques. For example, the MADM system or CLoNe (Garcia-Moreno et al., in vivo. Finally, we would like to emphasize that it is critical to consider an evolutionary perspective in order to understand brain development as a whole system.In conclusion, by using these imaging techniques in combination with conventional genetic manipulation, we could advance our understanding of the molecular mechanisms underlying neuronal migration and brain development The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Although vitamin A dimers are important physiological players in the visual cycle, their role in retinal degeneration is still poorly understood. To elucidate this, Ilyas Washington and colleagues investigated the effects of intravenous administration of A2E, obtained by a single-step chromatography-free method, in rabbits. The authors found that A2E induced photoreceptor damage together with inflammation and remodelling of blood vessels in the RPE and the supporting BM, leading to retinal degeneration. These results suggest that accumulation of vitamin A dimers is not a secondary symptom but rather a primary driver of retinal degeneration. Targeting vitamin A dimerisation might represent a strategy to prevent several forms of blindness.