Development of ISB 1442, a CD38 and CD47 bispecific biparatopic antibody innate cell modulator for the treatment of multiple myeloma.

Antibody engineering can tailor the design and activities of therapeutic antibodies for better efficiency or other advantageous clinical properties. Here we report the development of ISB 1442, a fully human bispecific antibody designed to re-establish synthetic immunity in CD38+ hematological malignancies. ISB 1442 consists of two anti-CD38 arms targeting two distinct epitopes that preferentially drive binding to tumor cells and enable avidity-induced blocking of proximal CD47 receptors on the same cell while preventing on-target off-tumor binding on healthy cells. The Fc portion of ISB 1442 is engineered to enhance complement dependent cytotoxicity, antibody dependent cell cytotoxicity and antibody dependent cell phagocytosis. ISB 1442 thus represents a CD47-BsAb combining biparatopic targeting of a tumor associated antigen with engineered enhancement of antibody effector function to overcome potential resistance mechanisms that hamper treatment of myeloma with monospecific anti-CD38 antibodies. ISB 1442 is currently in a Phase I clinical trial in relapsed refractory multiple myeloma.

[Impact of the 21-gene assay in decision-making during multidisciplinary breast meeting: A French experience]. / Impact d'un test de signature génomique en sénologie sur les décisions en réunion de concertation pluridisciplinaire : une expérience française.

OBJECTIVES: The 21-gene assay (Oncotype DX(®)) test is used to estimate the risk of recurrence and to predict the benefit of adjuvant chemotherapy at an early stage of endocrine responsive breast cancers, without HER2 overexpression or amplification. This test corresponds to a recurrence score (RS), classifying patients into three groups (low, intermediate or high risk). The objective of this two-center prospective study is to define the impact of Oncotype DX(®) in clinical practice. METHODS: Between August 2013 and May 2015, an Oncotype DX(®) test was decided in multidisciplinary meeting, to certain patients with an indication of adjuvant chemotherapy for HR+ and HER2 negative cancers. The therapeutic changes after knowledge of RS were collected. An estimate of the economic impact was performed and a correlation between the RS and usual breast cancer prognostic markers was investigated. RESULTS: Thirty-nine patients had a test, twenty-six (66.7%) of them have finally been no indication retaining chemotherapy. The economy obtained through the use of the test was estimated around 173,000euros. It has not been demonstrated correlation between the RS, the usual decisional and prognostic factors for breast cancer or with adjuvant! Online. CONCLUSIONS: The RS has an additional decision value compared to other common decision criteria. Use of Oncotype DX(®) reduced in our experience the indications of adjuvant chemotherapy. The medical and economic impact could be significant.

Observation of nanophase segregation in LiCl aqueous solutions from transient grating experiments.

Transient grating experiments performed on supercooled LiCl, RH2O solutions with R > 6 reveal the existence of well resolved, short time, extra signal which superposes to the normal signal observed for the R = 6 solution and for homogenous glass forming systems. This extra signal shows up below 190 K, its shape and the associated timescale depend only on temperature, while its intensity increases with R. We show that the origin of this signal is a phase separation between clusters with a low solute concentration and the remaining, more concentrated, solution. Our analysis demonstrates that these clusters have a nanometer size and a composition which are rather temperature independent, while increasing R simply increases the density of these clusters.

The low frequency dynamics of supercooled LiBr, 6H2O.

We present results of a series of experiments performed on LiBr, 6H(2)0 from room temperature down to 172 K ≈ 1.2T(g). These ultrasound, Brillouin and depolarized light scattering, and transient grating experiments show that, above 215 K, this solution behaves like supercooled water: its zero frequency sound velocity C(0) continuously decreases with decreasing temperature, and the reorientational dynamics of the water molecules can be directly detected at some temperatures of this domain. Conversely, below 215 K, a new regime sets in, where the apparent C(0) is practically temperature independent and where a ß, Arrenhius like, relaxation process coexists with the usual, Vogel-Fulcher like, α relaxation process of the supercooled liquid. These results are similar to those recently obtained in LiCl, 6H(2)O. The onset of the new regime is possibly due to an increase of the interaction of the water molecules with a neighboring Li(+) ion when lowering the temperature. We also compare our results with published dielectric data on water solutions of glass forming polyalcohols. Some of them present a low temperature splitting of their relaxation time similar to what is found in LiBr, 6H(2)O.

The low frequency phonons dynamics in supercooled LiCl, 6 H2O.

We report the results of a series of ultrasound, Brillouin scattering, and optical heterodyne detected transient grating experiments performed on a LiCl, 6H(2)O solution from room temperature down to the vicinity of its liquid-glass transition, T(g) approximately 138 K. Down to T approximately 215 K, the supercooled liquid has a behavior similar to what is expected for supercooled water: its zero frequency sound velocity, C(0), continuously decreases while the corresponding infinite frequency velocity, C(infinity), sharply increases, reflecting the increasing importance of H bonding when temperature is lowered. Below 215 K, specific aspects of the solution, presumably related to the role of the Li(+) and Cl(-) ions, modify the thermal behavior of C(0), while a beta relaxation process also appears and couples to the sound propagation. The origin of those two effects is briefly discussed.

Scaling of the structural relaxation in simulated liquid silica.

A scaling law for the alpha relaxation time tau , involving the ratio of a density-dependent energy to the thermal energy, has been found to hold in many fragile glass-forming liquids. This scaling has been recently linked to a local quantity n{loc}(rho,T) relating the variation of tau with density to its variation with temperature. In many fragile liquids, the variation of n{loc}(rho,T) is weak enough to take it as constant over the experimental temperature and density domain. We show that simulated liquid silica has an opposite behavior; close to T{g}, n{loc} is negative for moderate densities and exhibits a significant variation with rho and T, reaching positive values for higher temperature and/or densities. Moreover, those variations linearly correlate to a measure of the bonding properties of the liquid.

Analysis of a heterodyne-detected transient-grating experiment on a molecular supercooled liquid. II. Application to m-toluidine.

This paper reports the first detailed analysis of a transient grating (TG) experiment on a supercooled molecular liquid, m-toluidine, from 330K (1.75Tg) to 190K (1.01Tg) based on the theoretical model presented in Paper I of this series. This method allows one to give a precise description, over a wide dynamical range, of the different physical phenomena giving rise to the signals. Disentangling the isotropic and the anisotropic parts of the TG response, a careful fitting analysis yields detailed information on the rotation-translation coupling function. We also extract the structural relaxation times related to the "longitudinal" viscosity over almost 10 decades in time and the corresponding stretching coefficient. The value of some other parameters and information on their thermal behavior is also reported.

Analysis of a heterodyne-detected transient-grating experiment on a molecular supercooled liquid. I. Basic formulation of the problem.

We present the basic equations necessary to interpret heterodyne-detected transient-grating experiments performed on a supercooled liquid composed of anisotropic molecules. The final expressions are given under a form suitable for their direct application to a test case.

High frequency acoustic excitations in ordered diblock copolymer studied by inelastic x-ray scattering.

The phonon propagation in lamellar nanostructures formed via self-assembling of short styrene-b-isoprene (SI) as well as of its more incompatible styrene-b-(ethylene-alt-propylene) (SEP) counterpart was studied by inelastic x-ray scattering. Irrespective of the physical state of the block copolymers, a single acoustic phonon was observed in SI (ordered and disordered) and SEP (ordered). At GHz frequencies, inelastic light scattering from the same samples revealed very small dispersion in the sound phase velocity but a short phonon lifetime.

Temperature and pressure study of Brillouin transverse modes in the organic glass-forming liquid orthoterphenyl.

Transverse Brillouin spectra of orthoterphenyl are measured in the (250-305 K; 0.1-100 MPa) temperature-pressure range, which corresponds to the supercooled phase of this organic glass former. We show that the analysis of these spectra combined with an extrapolation of the reorientation times under pressure leads to an estimate of the static shear viscosity in a pressure range whose validity extends beyond the range of the Brillouin measurements. The relative contributions of temperature and of density to the change of this reorientation time measured along an isobar are extracted from our results in a large temperature range extending from the liquid to the low temperature supercooled state. They appear to be always of the same order of magnitude. It is also shown that in the range of the experiment, the orientational time is depending on a unique parameter built on temperature and density.

Differential regulation of p75 and trkB mRNA expression after depolarizing stimuli or BDNF treatment in basal forebrain neuron cultures.

Extensive evidence suggests that BDNF regulates neural function and architecture after depolarization. Expression of BDNF is increased after depolarization, and the ability of BDNF to modulate synaptic function is well documented. To further investigate BDNF signaling after activity, we analyzed the effects of depolarization or BDNF treatment on receptor mRNA expression in cultured basal forebrain neurons. Levels of mRNA coding for the cognate BDNF receptor, trkB, as well as the common neurotrophin receptor, p75, were quantitated simultaneously using a sensitive solution hybridization technique. Depolarization or BDNF treatment increased p75 mRNA expression 94% and 195%, respectively. In contrast, trkB message decreased 23% after depolarization but was unchanged by BDNF treatment. Together, these changes resulted in significant increases in the p75/trkB ratio after depolarization or BDNF treatment that could alter BDNF binding or signal transduction.

Neuronal signals regulate neurotrophin expression in oligodendrocytes of the basal forebrain.

Previous studies suggest that oligodendrocytes express trophic molecules, including neurotrophins. These molecules have been shown to influence nearby neurons. To determine whether neuronal signals may, in turn, affect oligodendrocyte-derived trophins, we examined regulation of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) mRNA expression in cultured oligodendrocytes of the basal forebrain. Neuronal signals had distinct effects on individual neurotrophins. KCl elicited increases in BDNF mRNA, but did not affect expression of NGF or NT-3. The cholinergic agonist, carbachol, increased expression of NGF, but did not affect expression of BDNF or NT-3. Glutamate elicited a decrease in BDNF, but did not affect expression of NGF or NT-3. This glutamate effect is not due to toxicity, since the number of total cells was unchanged, while the number of mature myelin basic protein positive (MBP+) cells increased. Our observations suggest that individual neuronal signals distinctly influence the trophic function of oligodendrocytes.

Brillouin scattering study of molten zinc chloride.

Polarized and depolarized Brillouin scattering experiments on molten ZnCl2 were performed between 300 and 600 degrees C in different geometries. VV spectra measured in backscattering and small angle scattering were analyzed with conventional viscoelastic theory using either a Debye or a Cole-Davidson model for the memory function. We also analyzed in the same way the temperature dependence of the transverse Brillouin lines detected in a 90 degrees VH geometry. We show that the Cole-Davidson memory function yields a consistent interpretation of all the spectra. The resulting shear and longitudinal relaxation times are equal within their error bars, and are about 2.5 times smaller than the alpha relaxation time previously determined. The static shear viscosity values deduced from the analysis of the propagating transverse waves agree, at all temperatures, with the measured viscosity values.

Brain-derived neurotrophic factor in astrocytes, oligodendrocytes, and microglia/macrophages after spinal cord injury.

Recent studies suggest that the injured adult spinal cord responds to brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT3) with enhanced neuron survival and axon regeneration. Potential neurotrophin sources and cellular localization in spinal cord are largely undefined. We examined glial BDNF localization in normal cord and its temporospatial distribution after injury in vivo. We used dual immunolabeling for BDNF and glial fibrillary acidic protein (GFAP) in astrocytes, adenomatous polyposis coli tumor suppressor protein (APC) for oligodendrocytes or type III CDH receptor (OX42) for microglia/macrophages. In normal cord, small subsets of astrocytes and microglia/macrophages and most oligodendrocytes exhibited BDNF-immunoreactivity. Following injury, the number of BDNF-immunopositive astrocytes and microglia/macrophages increased dramatically at the injury site over time. Most oligodendrocytes contained BDNF 1 day and 1 week following injury, but APC-positive cells were largely absent at the injury site 6 weeks postinjury. Glial BDNF-immunolabeling was also examined 10 and 20 mm from the wound. Ten millimeters from the lesion, astrocyte and microglia/macrophage BDNF-immunolabeling resembled that at the injury at all times examined. Twenty millimeters from injury, BDNF localization in all three glial subtypes resembled controls, regardless of time postlesion. Our findings suggest that in normal adult cord, astrocytes, oligodendrocytes, and microglia/macrophages play roles in local trophin availability and in trophin-mediated injury and healing responses directly within and surrounding the wound site.

Multiple ephrins regulate hippocampal neurite outgrowth.

Increasing evidence indicates that the eph family of ligands and receptors guides the formation of topographic maps in the brain through repulsive interactions. For example, we have recently found that in the hippocamposeptal system, the ligand ephrin-A2, which is expressed in an increasing gradient from dorsal to ventral septum, selectively induces pruning of topographically inappropriate medial hippocampal axons. The recent detection of ephrins A3 and A5, as well as A2, in the septum raised critical functional questions. Do the ligands act combinatorially, ensuring appropriate three-dimensional spatiotemporal projection, or do they exert entirely distinct actions in addition to guidance mechanisms? To approach these alternatives, we cloned mouse ephrin-A2 and compared the activities of the three ligands. Here, we show that these ligands reduce the number of hippocampal neurites in a similar fashion. The effect was regionally specific; medial hippocampal neurites were reduced 1.5- to 1.8-fold, whereas lateral hippocampal neurites were not significantly affected, conforming to topographic projection in vivo. Furthermore, we found that ephrins regulated neurite number in a stage-specific fashion, affecting E19 hippocampal neurites more than E16 neurites. Our observations suggest that all three septal ephrins, A2, A3, and A5, play spatiotemporally specific roles in guiding topographic projections from the hippocampus.

Expression of neurotrophins in the adult spinal cord in vivo.

Potential roles of trophins in the normal and injured spinal cord are largely undefined. However, a number of recent studies suggest that adult spinal cord expresses neurotrophin receptors and responds to the neurotrophins, brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT3), particularly after injury. The data indicate that trophins may enhance regrowth after damage and may represent a new therapeutic approach to injury. Neurotrophins are reportedly present in the spinal cord, but the cellular localization is unknown. This information is critical to begin delineating mechanisms of actions. To approach this problem, we examined whether spinal cord glia express BDNF and NT3 in vivo and have begun to define cellular distribution. Specific antibodies directed against the neurotrophins were utilized to visualize neurotrophin protein. Initial studies indicated that small cells in the white matter of adult rat spinal cord express BDNF and NT3. Large neurotrophin-positive neurons were also identified in the ventral cord. To identify the neurotrophin-positive cells, co-localization studies were performed utilizing neurotrophin polyclonal antisera together with monoclonal antibodies directed against the astrocyte marker, glial fibrillary acidic protein (GFAP). In the white matter of adult spinal cord, GFAP-positive and GFAP-negative cells expressed BDNF and NT3. Our study suggests that astrocyte and non-astrocyte cells provide trophic support to the adult spinal cord.

Ephrin-dependent growth and pruning of hippocampal axons.

Neuronal connections are arranged topographically such that the spatial organization of neurons is preserved by their termini in the targets. During the development of topographic projections, axons initially explore areas much wider than the final targets, and mistargeted axons are pruned later. The molecules regulating these processes are not known. We report here that the ligands of the Eph family tyrosine kinase receptors may regulate both the initial outgrowth and the subsequent pruning of axons. In the presence of ephrins, the outgrowth and branching of the receptor-positive hippocampal axons are enhanced. However, these axons are induced later to degenerate. These observations suggest that the ephrins and their receptors may regulate topographic map formation by stimulating axonal arborization and by pruning mistargeted axons.

Unilateral hippocampal lesions in newborn and adult rats: effects on spatial memory and BDNF gene expression.

Subcortical damage at birth often produces more severe deficits than similar lesions in an adult. In the present study, effects of unilateral electrolytic hippocampal ablations made on postnatal day 1 or in 3-month-old adult rats, were compared. Exploratory behavior and spatial navigation in the Morris water maze (MWM) were assessed 8 and 20 weeks after hippocampal damage. Rats with neonatal damage did not respond to novelty in the environment and did not learn to find the hidden platform in the MWM. Rats lesioned as adults did learn the water maze task, but slower than controls. We hypothesized that behavioral deficits observed in rats lesioned at birth, may be due, in part, to neurochemical dysfunction of the contralateral hippocampus. Specifically, cholinergic and GABAergic neurotransmission were assessed by measuring choline-acetyltransferase (ChAT) and GABAdecarboxylase (GAD) activity. In addition, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) mRNA levels were assayed in the remaining (contralateral) hippocampus. Of these molecules, only BDNF gene expression was significantly reduced (by 30%) at 8 and 20 weeks after neonatal and adult unilateral ablation. The similar reduction in BDNF mRNA in both treatment groups does not correspond with the lesion's differential effect on memory function. However, the more severe learning impairment after neonatal lesion may reflect increased dependence on trophins during development.

Differential involvement of metabotropic and p75 neurotrophin receptors in effects of nerve growth factor and neurotrophin-3 on cultured Purkinje cell survival.

We have examined the role of the p75 neurotrophin receptor in survival-promoting effects of nerve growth factor (NGF) and neurotrophin-3 (NT-3) on cultured Purkinje cells. Previously, we showed that NGF promotes Purkinje cell survival in conjunction with (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD), an agonist of metabotropic excitatory amino acid receptors, whereas NT-3 by itself increases cell number. We now present evidence that p75 plays different roles in Purkinje cell responses to the two neurotrophins. A metabotropic receptor of the mGluR1 subtype may interact with p75 function, so as to regulate Purkinje cell responsiveness to neurotrophins. When cerebellar cultures were grown for 6 days in the presence of ACPD and a mutant form of NGF that does not bind to p75, no increase in Purkinje cell number was observed. Moreover, the survival-promoting effect of wild-type NGF and ACPD could be inhibited by a neutralizing antiserum to p75 or by a pyrazoloquinazolinone inhibitor of neurotrophin binding to p75. In contrast, the response to NT-3 was potentiated by anti-p75 treatment and by the quinazolinone. These data indicate the mediation of p75 in the trophic response to NGF-ACPD and a negative modulatory role of p75 in the action of NT-3. To probe the role of ACPD in the p75-dependent response to NGF, metabotropic receptor subtype-specific ligands were tested. The pattern of agonist specificity implicated the mGluR1 subtype, a receptor that is expressed at high levels by Purkinje cells and linked to activation of protein kinase C (PKC). Down-regulation or blockade of PKC abolished the response to NGF-ACPD. Consistent with the opposite roles of p75 in effects of the two neurotrophins, blockade of mGluR1 or PKC potentiated the survival response elicited by NT-3. In sum, our data suggest that afferent excitatory transmitters activate specific metabotropic receptors to elicit a p75-mediated action of NGF. NT-3 acts on Purkinje cells by a different mechanism that is not absolutely p75-dependent and that is reduced by neurotrophin access to p75 and metabotropic receptor activity.

Neurotransmitters and neurotrophins collaborate to influence brain development.

It is well recognized that the neurotrophin family of factors as well as neurotransmitters play critical roles in the ontogeny of the brain. Moreover, a growing literature suggests that these environmental signals do not operate individually, but interact in critical ways to enhance maturation. This review focuses on three brain systems where this collaboration is particularly evident: the cerebellum, the basal forebrain-hippocampus and locus coeruleus-hippocampus. The material presented indicates that cross-talk between neurotransmitters and neurotrophins may be a mechanism common to the development of multiple neuronal groups throughout the central nervous system. Moreover, this cross-talk appears to involve the interaction of both neuronal and glial cell populations.