Resistance to mTORC1 Inhibitors in Cancer Therapy: From Kinase Mutations to Intratumoral Heterogeneity of Kinase Activity.

Targeting mTORC1 has been thoroughly explored in cancer therapy. Following encouraging preclinical studies, mTORC1 inhibitors however failed to provide substantial benefits in cancer patients. Several resistance mechanisms have been identified including mutations of mTOR and activation of alternate proliferation pathways. Moreover, emerging evidence discloses intratumoral heterogeneity of mTORC1 activity that further contributes to a reduced anticancer efficacy of mTORC1 inhibitors. Genetic heterogeneity as well as heterogeneous conditions of the tumor environment such as hypoxia profoundly modifies mTORC1 activity in tumors and hence influences the response of tumors to mTORC1 inhibitors. Intriguingly, the heterogeneity of mTORC1 activity also occurs towards its substrates at the single cell level, as mutually exclusive pattern of activation of mTORC1 downstream effectors has been reported in tumors. After briefly describing mTORC1 biology and the use of mTORC1 inhibitors in patients, this review will give an overview on concepts of resistance to mTORC1 inhibition in cancer with a particular focus on intratumoral heterogeneity of mTORC1 activity

Differential interferometric phases at high spectral resolution as a sensitive physical diagnostic of circumstellar disks

Context. The circumstellar disks ejected by many rapidly rotating B stars (so-called Be stars) offer the rare opportunity of studying the structure and dynamics of gaseous disks at high spectral as well as angular resolution. Aims. This paper explores a newly identified effect in spectro-interferometric phase that can be used for probing the inner regions of gaseous edge-on disks on a scale of a few stellar radii. Methods. The origin of this effect (dubbed central quasi-emission phase signature, CQE-PS) lies in the velocity-dependent line absorption of photospheric radiation by the circumstellar disk. At high spectral and marginal interferometric resolution, photocenter displacements between star and isovelocity regions in the Keplerian disk reveal themselves through small interferometric phase shifts. To investigate the diagnostic potential of this effect, a series of models are presented, based on detailed radiative transfer calculations in a viscous decretion disk. Results. Amplitude and detailed shape of the CQE-PS depend sensitively on disk density and size and on the radial distribution of the material with characteristic shapes in differential phase diagrams. In addition, useful lower limits to the angular size of the central stars can be derived even when the system is almost unresolved. Conclusions. The full power of this diagnostic tool can be expected if it can be applied to observations over a full life-cycle of a disk from first ejection through final dispersal, over a full cycle of disk oscillations, or over a full orbital period in a binary system

Fine-Tuning Tumor Endothelial Cells to Selectively Kill Cancer.

Tumor endothelial cells regulate several aspects of tumor biology, from delivering oxygen and nutrients to shaping the immune response against a tumor and providing a barrier against tumor cell dissemination. Accordingly, targeting tumor endothelial cells represents an important modality in cancer therapy. Whereas initial anti-angiogenic treatments focused mainly on blocking the formation of new blood vessels in cancer, emerging strategies are specifically influencing certain aspects of tumor endothelial cells. For instance, efforts are generated to normalize tumor blood vessels in order to improve tumor perfusion and ameliorate the outcome of chemo-, radio-, and immunotherapy. In addition, treatment options that enhance the properties of tumor blood vessels that support a host's anti-tumor immune response are being explored. Hence, upcoming anti-angiogenic strategies will shape some specific aspects of the tumor blood vessels that are no longer limited to abrogating angiogenesis. In this review, we enumerate approaches that target tumor endothelial cells to provide anti-cancer benefits and discuss their therapeutic potential

Revealing the structure of the outer disks of Be stars

Context. The structure of the inner parts of Be star disks (20 stellar radii) is well explained by the viscous decretion disk (VDD) model, which is able to reproduce the observable properties of most of the objects studied so far. The outer parts, on the ther hand, are not observationally well-explored, as they are observable only at radio wavelengths. A steepening of the spectral slope somewhere between infrared and radio wavelengths was reported for several Be stars that were previously detected in the radio, but a convincing physical explanation for this trend has not yet been provided. Aims. We test the VDD model predictions for the extended parts of a sample of six Be disks that have been observed in the radio to address the question of whether the observed turndown in the spectral energy distribution (SED) can be explained in the framework of the VDD model, including recent theoretical development for truncated Be disks in binary systems. Methods. We combine new multi-wavelength radio observations from the Karl. G. Jansky Very Large Array (JVLA) and Atacama Pathfinder Experiment (APEX) with previously published radio data and archival SED measurements at ultraviolet, visual, and infrared wavelengths. The density structure of the disks, including their outer parts, is constrained by radiative transfer modeling of the observed spectrum using VDD model predictions. In the VDD model we include the presumed effects of possible tidal influence from faint binary companions. Results. For 5 out of 6 studied stars, the observed SED shows strong signs of SED turndown between far-IR and radio wavelengths. A VDD model that extends to large distances closely reproduces the observed SEDs up to far IR wavelengths, but fails to reproduce the radio SED. ... (abstract continues but did not fit here)Comment: 20 pages, 8 figure

Stellar parameters of Be stars observed with X-shooter

Aims. The X-shooter archive of several thousand telluric star spectra was skimmed for Be and Be-shell stars to derive the stellar fundamental parameters and statistical properties, in particular for the less investigated late type Be stars, and the extension of the Be phenomenon into early A stars. Methods. An adapted version of the BCD method is used, utilizing the Balmer discontinuity parameters to determine effective temperature and surface gravity. This method is optimally suited for late B stars. The projected rotational velocity was obtained by profile fitting to the Mg ii lines of the targets, and the spectra were inspected visually for the presence of peculiar features such as the infrared Ca ii triplet or the presence of a double Balmer discontinuity. The Balmer line equivalent widths were measured, but due to uncertainties in determining the photospheric contribution are useful only in a subsample of Be stars for determining the pure emission contribution. Results. A total of 78 Be stars, mostly late type ones, were identified in the X-shooter telluric standard star archive, out of which 48 had not been reported before. The general trend of late type Be stars having more tenuous disks and being less variable than early type ones is confirmed. The relatively large number (48) of relatively bright (V > 8.5) additional Be stars casts some doubt on the statistics of late type Be stars; they are more common than currently thought: The Be/B star fraction may not strongly depend on spectral subtype.Comment: Accepted for publication in A&

Multitechnique testing of the viscous decretion disk model I. The stable and tenuous disk of the late-type Be star β\beta CMi

The viscous decretion disk (VDD) model is able to explain most of the currently observable properties of the circumstellar disks of Be stars. However, more stringent tests, focusing on reproducing multitechnique observations of individual targets via physical modeling, are needed to study the predictions of the VDD model under specific circumstances. In the case of nearby, bright Be star $\beta$ CMi, these circumstances are a very stable low-density disk and a late-type (B8Ve) central star. The aim is to test the VDD model thoroughly, exploiting the full diagnostic potential of individual types of observations, in particular, to constrain the poorly known structure of the outer disk if possible, and to test truncation effects caused by a possible binary companion using radio observations. We use the Monte Carlo radiative transfer code HDUST to produce model observables, which we compare with a very large set of multitechnique and multiwavelength observations that include ultraviolet and optical spectra, photometry covering the interval between optical and radio wavelengths, optical polarimetry, and optical and near-IR (spectro)interferometry. Due to the absence of large scale variability, data from different epochs can be combined into a single dataset. A parametric VDD model with radial density exponent of $n$ = 3.5, which is the canonical value for isothermal flaring disks, is found to explain observables typically formed in the inner disk, while observables originating in the more extended parts favor a shallower, $n$ = 3.0, density falloff. Modeling of radio observations allowed for the first determination of the physical extent of a Be disk (35$^{+10}_{-5}$ stellar radii), which might be caused by a binary companion. Finally, polarization data allowed for an indirect measurement of the rotation rate of the star, which was found to be $W \gtrsim 0.98$, i.e., very close to critical.Comment: 19 pages (35 including online material), 17 figures, 2 online figures, 2 online tables with dat

Acidic pH reduces VEGF-mediated endothelial cell responses by downregulation of VEGFR-2; relevance for anti-angiogenic therapies.

Anti-angiogenic treatments targeting the vascular endothelial growth factor or its receptors have shown clinical benefits. However, impact on long-term survival remains limited. Solid tumors display an acidic microenvironment that profoundly influences their biology. Consequences of acidity on endothelial cells and anti-angiogenic therapies remain poorly characterized and hence are the focus of this study. We found that exposing endothelial cells to acidic extracellular pH resulted in reduced cell proliferation and migration. Also, whereas VEGF increased endothelial cell proliferation and survival at pH 7.4, it had no effect at pH 6.4. Furthermore, in acidic conditions, stimulation of endothelial cells with VEGF did not result in activation of downstream signaling pathways such as AKT. At a molecular level, acidity significantly decreased the expression of VEGFR-2 by endothelial cells. Consequently, anti-angiogenic therapies that target VEGFR-2 such as sunitinib and sorafenib failed to block endothelial cell proliferation in acidic conditions. In vivo, neutralizing tumor acidity with sodium bicarbonate increased the percentage of endothelial cells expressing VEGFR-2 in tumor xenografts. Furthermore, combining sodium bicarbonate with sunitinib provided stronger anti-cancer activity than either treatment alone. Histological analysis showed that sunitinib had a stronger anti-angiogenic effect when combined with sodium bicarbonate. Overall, our results show that endothelial cells prosper independently of VEGF in acidic conditions partly as a consequence of decreased VEGFR-2 expression. They further suggest that strategies aiming to raise intratumoral pH can improve the efficacy of anti-VEGF treatments

Acidic tumor microenvironment abrogates the efficacy of mTORC1 inhibitors.

Blocking the mechanistic target of rapamycin complex-1 (mTORC1) with chemical inhibitors such as rapamycin has shown limited clinical efficacy in cancer. The tumor microenvironment is characterized by an acidic pH which interferes with cancer therapies. The consequences of acidity on the anti-cancer efficacy of mTORC1 inhibitors have not been characterized and are thus the focus of our study. Cancer cell lines were treated with rapamycin in acidic or physiological conditions and cell proliferation was investigated. The effect of acidity on mTORC1 activity was determined by Western blot. The anticancer efficacy of rapamycin in combination with sodium bicarbonate to increase the intratumoral pH was tested in two different mouse models and compared to rapamycin treatment alone. Histological analysis was performed on tumor samples to evaluate proliferation, apoptosis and necrosis. Exposing cancer cells to acidic pH in vitro significantly reduced the anti-proliferative effect of rapamycin. At the molecular level, acidity significantly decreased mTORC1 activity, suggesting that cancer cell proliferation is independent of mTORC1 in acidic conditions. In contrast, the activation of mitogen-activated protein kinase (MAPK) or AKT were not affected by acidity, and blocking MAPK or AKT with a chemical inhibitor maintained an anti-proliferative effect at low pH. In tumor mouse models, the use of sodium bicarbonate increased mTORC1 activity in cancer cells and potentiated the anti-cancer efficacy of rapamycin. Combining sodium bicarbonate with rapamycin resulted in increased tumor necrosis, increased cancer cell apoptosis and decreased cancer cell proliferation as compared to single treatment. Taken together, these results emphasize the inefficacy of mTORC1 inhibitors in acidic conditions. They further highlight the potential of combining sodium bicarbonate with mTORC1 inhibitors to improve their anti-tumoral efficacy

Targeting carbonic anhydrase IX improves the anti-cancer efficacy of mTOR inhibitors.

The inhibition of the mechanistic target of rapamycin complex 1 (mTORC1) by chemical inhibitors, such as rapamycin, has demonstrated anti-cancer activity in preclinical and clinical trials. Their efficacy is, however, limited and tumors eventually relapse through resistance formation. In this study, using two different cancer mouse models, we identify tumor hypoxia as a novel mechanism of resistance of cancer cells against mTORC1 inhibitors. Indeed, we show that the activity of mTORC1 is mainly restricted to the non-hypoxic tumor compartment, as evidenced by a mutually exclusive staining pattern of the mTORC1 activity marker pS6 and the hypoxia marker pimonidazole. Consequently, whereas rapamycin reduces cancer cell proliferation in non-hypoxic regions, it has no effect in hypoxic areas, suggesting that cancer cells proliferate independently of mTORC1 under hypoxia. Targeting the hypoxic tumor compartment by knockdown of carbonic anhydrase IX (CAIX) using short hairpin RNA or by chemical inhibition of CAIX with acetazolamide potentiates the anti-cancer activity of rapamycin. Taken together, these data emphasize that hypoxia impairs the anti-cancer efficacy of rapalogs. Therapeutic strategies targeting the hypoxic tumor compartment, such as the inhibition of CAIX, potentiate the efficacy of rapamycin and warrant further clinical evaluation