Unfinished business: Incomplete laminin processing exposes a tumor target

The G45 domain of laminin-332 is cleaved in normal tissues but remains in squamous cell carcinomas. Targeting this domain using a G45 antibody reduced in vivo tumor growth and invasion and suggests a role for G45 as a new therapeutic target.     

Unfinished business: Incomplete laminin processing exposes a tumor target

The G45 domain of laminin-332 is cleaved in normal tissues but remains in squamous cell carcinomas. Targeting this domain using a G45 antibody reduced in vivo tumor growth and invasion and suggests a role for G45 as a new therapeutic target.Key words: laminin, extracellular matrix, cancer, squamous cell carcinoma, invasionLaminin-332 (previously known as laminin-5) is a trimeric extracellular glycoprotein important for the integrity of the basement membrane zone.¹ It is found at high levels in squamous cell carcinomas (SCC) and its expression correlates with increased tumor invasiveness and poor prognosis.² In a recent article in Cancer Research, Marinkovich and colleagues³ show that the G45 domain of laminin-332, normally removed by proteolytic processing, remains present in SCC tissue.To determine the function of the G45 domain, the authors expressed the wild-type α3 chain of laminin-332 or the α3 chain lacking G45 (ΔG45) in normal and transformed laminin-332-null keratinocytes. They then examined whether the phenotype of the G45 deletion mutant could be reverted by re-expressing the G45 domain peptide.When expressed in laminin-332-null keratinocytes, the wild-type form of the protein was found deposited in the extracellular matrix. Reduced matrix deposition was observed with the ΔG45 mutant. In addition, the ΔG45 mutant was associated with abnormal peripheral cell adhesions, in contrast to central α6 integrin-associated adhesions in the wild-type. ΔG45 cells detached more rapidly in trypsin and showed increased cell migration in vitro. Expression of the G45 domain peptide restored laminin-332 deposition and reversed the changes in cell adhesion and migration.After transformation with Ras and IκBα, laminin-null keratinocytes are tumorigenic and invade Matrigel. Expression of wild-type laminin-332 increases invasiveness in these cells. Invasion was also increased, but to a lesser degree, with the ΔG45 mutant expressing cells. ΔG45 cells had reduced MMP levels, and both invasion and MMP production were restored by expression of the G45 domain. Similar results were seen in an in vivo SCC model where reduced tumor growth and muscle invasion correlated with expression of the ΔG45 mutant relative to wild-type protein. Interestingly, activation of the PI3K pathway almost completely restored tumor growth and laminin-332 deposition in the ΔG45 mutant expressing cells. In vitro, ΔG45 cells show decreased Akt phosphorylation and phospho- ERK nuclear translocation. Both could be corrected by activating PI3K. Together, these results suggest that G45 may promote laminin- 332 deposition via a signaling mechanism rather than acting simply as a matrix anchor.As G45 promotes tumorigenesis in SCC and is absent in normal tissues, could it be a target for anti cancer therapy in vivo? The authors show that an antibody targeting G45 dramatically inhibited subcutaneous SCC tumor growth. Intriguingly, despite recognizing both G45 and native laminin-332 by western blot, the antibody caused no disruption of epithelial-mesenchymal integrity in normal tissues.This study has shown an important role for the G45 domain of laminin-332 in SCC tumorigenesis. Considering its absence in normal tissues, G45 represents a potential target for anti-cancer therapy in human SCC.     

Unfinished business: efforts to define dual-use research of bioterrorism concern

Biotechnological research poses a special security problem because of the duality between beneficial use and misuse. In order to find a balance between regulating potentially dangerous research and assuring scientific advancement, a number of assessments have tried to define which types of research are especially open to misuse and should therefore be considered dual-use research of special concern requiring rigorous oversight. So far, there has been no common understanding of what such activities are. Here we present a review of 27 assessments focusing on biological dual-use issues published between 1997 and 2008. Dual-use research activities identified by these assessments as being of special concern were compiled and compared. Moreover, from these 27 assessments, the primary research publications explicitly identified as examples of concerning research activities were extracted and analyzed. We extracted a core list of 11 activities of special concern and show that this list does not match with the reasons why primary research publications were identified as being of special concern. Additionally, we note that the 11 activities identified are not easily conducted or replicated, and therefore the likelihood of their being used in a high-tech mass casualty bioterrorism event should be reevaluated.     

Paramutation: an encounter leaving a lasting impression

Paramutation is the result of heritable changes in gene expression that occur upon interaction between alleles. Whereas Mendelian rules, together with the concept of genetic transmission via the DNA sequence, can account for most inheritance in sexually propagating organisms, paramutation-like phenomena challenge the exclusiveness of Mendelian inheritance. Most paramutation-like phenomena have been observed in plants but there is increasing evidence for its occurrence in other organisms, including mammals. Our knowledge of the underlying mechanisms, which might involve RNA silencing, physical pairing of homologous chromosomal regions or both, is still limited. Here, we discuss the characteristics of different paramutation-like interactions in the light of arguments supporting each of these alternative mechanisms.     

Mechanisms of stimulation of olfactory neurons: an essay

It is widely believed that the mechanism by which olfactoryneurons are stimulated by odorants is via specific receptorproteins located on or in the apical cell membranes of the neurons.In this review alternative mechanisms are explored and a modelis presented in which sensitivity and specificity can be accountedfor by the general irritability of cells to chemicals. The premiseson which the model is based are: (i) Groups of olfactory nerves fire action potentials whichare summated at the first synapse, in the olfactory bulb. Theseneurons fire spontaneously because their resting potentialsare unstable. (ii) What we call olfactory quality is encoded in patterns offiring of different olfactory neurons. (iii) Olfactory neurons are not identical. The sources of theirindividuality include differences in their age, spatial locationin the plane of the tissue, depth within the tissue and distancefrom the nearest capillary. The heterogeneity will be reflectedas differences in their metabolism and chemical composition. (iv) Most of the surface area of olfactory neurons is bathedin mucus that is secreted by Bowman's glands and sustentacularcells. Therefore, the resting potential and firing frequencyof each will depend upon the composition of the mucus. It follows from these premises that alterations in the mucuscomposition, extracellular fluid composition, metabolic stateor properties of neuronal membranes will alter the resting potentialand, therefore, the firing frequency of each neuron. The effectswill vary among neurons because of their individuality evenif the stimulus is distributed uniformly across the olfactorymucosa, different regions will be exposed to different patternsof stimulus concentration because odorants are differentiallyadsorbed and metabolized. According to this scheme qualitativelydifferent patterns of neural activity will result from differentodorants, the firing of some neurons being inhibited and thatof others being stimulated in each case. This will be true independentlyof the existence of specific receptor proteins, the effectsof which must be superimposed upon the general effects of nonspecificirritability.     

A jury of none: an essay on the last acceptable form of civic banishment

Though many socio-legal scholars have criticized those measures that deny convicted felons the right to vote, few have challenged the statutes that withhold a convicted felon’s opportunity to sit on a jury. A majority of US jurisdictions bar felons from jury service permanently, creating a class of citizens defined and punished by the criminal justice system but unable to impact its function. This essay considers the invisibility of felon jury exclusion, notes its unlikely supporters, and asserts that by preventing convicted felons from taking part in civic processes, authorities work against reentry initiatives.     

Knowing human moral knowledge to be true: an essay on intellectual conviction

The question addressed in this article is how people come to know the foundational axioms of their moral systems as true and correct. Drawing on my fieldwork among the Himba of northwestern Namibia, I argue that the most potent form of intellectual conviction is not generated through the external manipulations of ritual, but through a deeply internal experience in which moral knowledge coalesces with a subjectively perceived experience of timeless universality.     

Patch leaving strategies and superparasitism: an asymmetric generalized war of attrition

When several competitors deplete a patch, it can be advantageous for each of them to stay provided that others leave, whereas, on the other hand, staying longer decreases the expected payoff for everyone. This situation can be considered as a generalized war of attrition. Previous studies have shown that optimal patch leaving strategies become stochastic and the expected leaving time is much larger than predicted by the marginal value theorem when competitors interfere. The possibility of superparasitism, as occurs for example in parasitoids, induces such interference. In addition, it gives several complications. First, the payoff of females that have left the patch is affected by the ovipositions of the remaining individuals. Second, differences in the arrival time of females cause payoff-relevant asymmetries, since females that arrived early on have parasitized more hosts in a patch at the moment superparasitism starts than those that arrived later. We show that this can be modelled as an asymmetric generalized war of attrition, and derive global characteristics of the ESS for simultaneous decisions on when to start superparasitism and when to leave a patch.     

Transductions to generate plant form and pattern: an essay on cause and effect

Many complex processes can be broken into transduction steps where one state is converted to another by a well defined activity. One difficulty for analysis is that transductions occur in chains or networks. Another, of primary concern here, is that a single transduction can be complex. Some such transductions can efficiently explain phenomena often thought to be summations or orchestrations of many simple transductions. Pattern formation is in this category. For a wide range of transductions one can define cause and effect in a differential equation. In its integral one can define the before and after states. The main experimental tactic to characterize unknown transductions is co-variation. The before state (input) is altered, change in the after state (output) is assayed. Thus an unknown transduction, with cause and effect embodied in the differential, is investigated through long-term changes in its integral. This is fully practical when all of the integral is known or readily surmised, as in simple discrete biochemical transductions. As causal differential expressions become complex, their integrals become more versatile in generating output because this changes not only with variation in the expression itself but also with boundary conditions and limits. These very features, however, make such a function increasingly intractable to discovery by co-variation. Only a small part of the integral is embodied in the before and after states; the remainder is not readily surmised. Accordingly, in contrast to reliance on the role of controls to deduce unknown simple transductions, the complex ones are generally established through formalization of the differential nature of the process itself.     

Ethylene signaling: the MAPK module has finally landed

Ethylene hormone responses are negatively regulated by the CTR1 protein, which has similarity to mitogen-activated protein kinase kinase kinases (MAPKKKs). Because of this similarity, it has long been speculated that ethylene signal transduction involves a MAPK cascade. Now, a recent paper provides compelling evidence for an ethylene-activated MAPK pathway. The implication is that CTR1 and the newly identified MAPKK and MAPKs comprise a MAPK module that regulates ethylene responses in plants.     

Gene therapy: light is finally in the tunnel

After two decades of ups and downs, gene therapy has recently achieved a milestone in treating patients with Leber’s congenital amaurosis (LCA). LCA is a group of inherited blinding diseases with retinal degeneration and severe vision loss in early infancy. Mutations in several genes, including RPE65, cause the disease. Using adeno-associated virus as a vector, three independent teams of investigators have recently shown that RPE65 can be delivered to retinal pigment epithelial cells of LCA patients by subretinal injections resulting in clinical benefits without side effects. However, considering the whole field of gene therapy, there are still major obstacles to clinical applications for other diseases. These obstacles include innate and immune barriers to vector delivery, toxicity of vectors and the lack of sustained therapeutic gene expression. Therefore, new strategies are needed to overcome these hurdles for achieving safe and effective gene therapy. In this article, we shall review the major advancements over the past two decades and, using lung gene therapy as an example, discuss the current obstacles and possible solutions to provide a roadmap for future gene therapy research.     

Pollinator-dependent crops: an increasingly risky business

Three-quarters of leading global food crops rely on animal pollination. With both managed and wild pollinators declining, is there reason for concern? Researchers are beginning to pin down the possible long-term risks.     

From the laboratory bench to the patient's bedside: an update on clinical trials with mesenchymal stem cells

Mesenchymal Stem Cells (MSCs) are non-hematopoietic multi-potent stem-like cells that are capable of differentiating into both mesenchymal and non-mesenchymal lineages. In fact, in addition to bone, cartilage, fat, and myoblasts, it has been demonstrated that MSCs are capable of differentiating into neurons and astrocytes in vitro and in vivo. MSCs are of interest because they are isolated from a small aspirate of bone marrow and can be easily expanded in vitro. As such, these cells are currently being tested for their potential use in cell and gene therapy for a number of human diseases. Nevertheless, there are still some open questions about origin, multipotentiality, and anatomical localization of MSCs. In this review, we discuss clinical trials based on the use of MSCs in cardiovascular diseases, such as treatment of acute myocardial infarction, endstage ischemic heart disease, or prevention of vascular restenosis through stem cell-mediated injury repair. We analyze data from clinical trials for treatment of osteogenesis imperfecta (OI), which is a genetic disease characterized by production of defective type I collagen. We describe progress for neurological disease treatment with MSC transplants. We discuss data on amyotrophic lateral sclerosis (ALS) and on lysosomal storage diseases (Hurler syndrome and metachromatic leukodystrophy). A section of review is dedicated to ongoing clinical trials, involving MSCs in treatment of steroid refractory Graft Versus Host Disease (GVHD); periodontitis, which is a chronic disease affecting periodontium and causing destruction of attachment apparatus, heart failure, and bone fractures. Finally, we will provide information about biotech companies developing MSC therapy.