Differential effects of selective inhibitors targeting the PI3K/AKT/mTOR pathway in acute lymphoblastic leukemia

Purpose: Aberrant PI3K/AKT/mTOR signaling has been linked to oncogenesis and therapy resistance in various malignancies including leukemias. In Philadelphia chromosome (Ph) positive leukemias, activation of PI3K by dysregulated BCR-ABL tyrosine kinase (TK) contributes to the pathogenesis and development of resistance to ABL-TK inhibitors (TKI). The PI3K pathway thus is an attractive therapeutic target in BCR-ABL positive leukemias, but its role in BCR-ABL negative ALL is conjectural. Moreover, the functional contribution of individual components of the PI3K pathway in ALL has not been established. Experimental design: We compared the activity of the ATP-competitive pan-PI3K inhibitor NVP-BKM120, the allosteric mTORC1 inhibitor RAD001, the ATP-competitive dual PI3K/mTORC1/C2 inhibitors NVP-BEZ235 and NVP-BGT226 and the combined mTORC1 and mTORC2 inhibitors Torin 1, PP242 and KU-0063794 using long-term cultures of ALL cells (ALL-LTC) from patients with B-precursor ALL that expressed the BCR-ABL or TEL-ABL oncoproteins or were BCR-ABL negative. Results: Dual PI3K/mTOR inhibitors profoundly inhibited growth and survival of ALL cells irrespective of their genetic subtype and their responsiveness to ABL-TKI. Combined suppression of PI3K, mTORC1 and mTORC2 displayed greater antileukemic activity than selective inhibitors of PI3K, mTORC1 or mTORC1 and mTORC2. Conclusions: Inhibition of the PI3K/mTOR pathway is a promising therapeutic approach in patients with ALL. Greater antileukemic activity of dual PI3K/mTORC1/C2 inhibitors appears to be due to the redundant function of PI3K and mTOR. Clinical trials examining dual PI3K/mTORC1/C2 inhibitors in patients with B-precursor ALL are warranted, and should not be restricted to particular genetic subtypes

Discovery of Small-Molecule Stabilizers of 14-3-3 Protein-Protein Interactions via Dynamic Combinatorial Chemistry

Protein-protein interactions (PPIs) play an important role in numerous biological processes such as cell-cycle regulation and multiple diseases. The family of 14-3-3 proteins is an attractive target as they serve as binding partner to various proteins and are therefore capable of regulating their biological activities. Discovering small-molecule modulators, in particular stabilizers, of such complexes via traditional screening approaches is a challenging task. Herein, we pioneered the first application of dynamic combinatorial chemistry (DCC) to a PPI target, to find modulators of 14-3-3 proteins. Evaluation of the amplified hits from the DCC experiments for their binding affinity via surface plasmon resonance (SPR), revealed that the low-micromolar (KD 15-16 μM) acylhydrazones are 14-3-3/synaptopodin PPI stabilizers. Thus, DCC appears to be ideally suited for the discovery of not only modulators but even the more elusive stabilizers of notoriously challenging PPIs

Blinatumomab compared with standard of care for the treatment of adult patients with relapsed/refractory Philadelphia chromosome–positive B-precursor acute lymphoblastic leukemia

Background: A single-arm, phase 2 trial demonstrated the efficacy and safety of blinatumomab, a bispecific T-cell\u2013engaging antibody construct, in patients with relapsed/refractory (r/r) Philadelphia chromosome\u2013positive (Ph+) acute lymphoblastic leukemia (ALL), a rare hematologic malignancy with limited treatment options. This study compared outcomes with blinatumomab with those of a historical control treated with the standard of care (SOC). Methods: The blinatumomab trial enrolled adult patients with Ph+ ALL who were r/r to at least 1 second-generation tyrosine kinase inhibitor (n = 45). Propensity score analysis (PSA) was used to compare outcomes with blinatumomab with those of an external cohort of similar patients receiving SOC chemotherapy (n = 55). The PSA mitigated confounding variables between studies by adjusting for imbalances in the age at diagnosis and start of treatment, sex, duration from diagnosis to most recent treatment, prior allogeneic hematopoietic stem cell transplantation, prior salvage therapy, and number of salvage therapies. Bayesian data augmentation was applied to improve power to 80% with data from a phase 3 blinatumomab study in r/r Philadelphia chromosome\u2013negative ALL. Results: In the PSA, the rate of complete remission or complete remission with partial hematologic recovery was 36% for blinatumomab and 25% for SOC, and this resulted in an odds ratio of 1.54 (95% confidence interval [CI], 0.61-3.89) or 1.70 (95% credible interval [CrI], 0.94-2.94) with Bayesian data augmentation. Overall survival favored blinatumomab over SOC, with a hazard ratio of 0.81 (95% CI, 0.57-1.14) or 0.77 (95% CrI, 0.61-0.96) with Bayesian data augmentation. Conclusions: These results further support blinatumomab as a treatment option for patients with r/r Ph+ ALL

From the Heart of The Ghoul: C and N Abundances in the Corona of Algol B

Chandra Low Energy Transmission Grating Spectrograph observations of Algol have been used to determine the abundances of C and N in the secondary star for the first time. The analysis was performed relative to similar observations of an adopted "standard" star HR 1099. It is demonstrated that HR 1099 and Algol are coronal twins in many respects and that their X-ray spectra are very similar in nearly all details, except for the observed strengths of C and N lines. The H-like transitions of C and N in the coronae of Algol and HR 1099 demonstrate that the surface abundances of Algol B have been strongly modified by CN-processing, as shown earlier by Schmitt & Ness (2002). It is found that N is enhanced in Algol B by a factor of 3 compared to HR 1099. No C lines are detected in the Algol spectrum, indicating a C depletion relative to HR 1099 by a factor of 10 or more. These C and N abundances indicate that Algol B must have lost at least half of its initial mass, and are consistent with predictions of evolutionary models that include non-conservative mass transfer and angular momentum loss through magnetic activity. Little or no dredge-up of material subjected to CN-processing has occurred on the subgiant component of HR 1099. It is concluded that Fe is very likely depleted in the coronae of both Algol and HR 1099 relative to their photospheres by 0.5 dex, and C, N and O by 0.3 dex. Instead, Ne is enhanced by up to 0.5 dex.Comment: 17 pages, 4 figures, ApJ accepte

The Coronae of AR Lac

We observed the coronally active eclipsing binary, AR Lac, with the High Energy Transmission Grating on Chandra for a total of 97 ks, spaced over five orbits, at quadratures and conjunctions. Contemporaneous and simultaneous EUV spectra and photometry were also obtained with the Extreme Ultraviolet Explorer. Significant variability in both X-ray and EUV fluxes were observed, dominated by at least one X-ray flare and one EUV flare. We saw no evidence of primary or secondary eclipses. X-ray flux modulation was largest at high temperature, indicative of flare heating of coronal plasma. Line widths interpreted in terms of Doppler broadening suggest that both binary stellar components are active. From line fluxes obtained from total integrated spectra, we have modeled the emission measure and abundance distributions. A strong maximum was found in the differential emission measure, characterized by peaks at log T = 6.9 and 7.4, together with a weak but significant cooler maximum near log T=6.2, and a moderately strong hot tail from log T= 7.6-8.2. Coronal abundances have a broad distribution and show no simple correlation with first ionization potential. While the resulting model spectrum generally agrees very well with the observed spectrum, there are some significant discrepancies, especially among the many Fe L-lines. Both the emission measure and abundance distributions are qualitatively similar to prior determinations from other X-ray and ultraviolet spectra, indicating some long-term stability in the overall coronal structure.Comment: 31 pages, 8 figures, 3 tables; Accepted for publication in the Astrophysical Journal (tentatively October 1, 2003

Continuous heating of a giant X-ray flare on Algol

Giant flares can release large amounts of energy within a few days: X-ray emission alone can be up to ten percent of the star's bolometric luminosity. These flares exceed the luminosities of the largest solar flares by many orders of magnitude, which suggests that the underlying physical mechanisms supplying the energy are different from those on the Sun. Magnetic coupling between the components in a binary system or between a young star and an accretion disk has been proposed as a prerequisite for giant flares. Here we report X-ray observations of a giant flare on Algol B, a giant star in an eclipsing binary system. We observed a total X-ray eclipse of the flare, which demonstrates that the plasma was confined to Algol B, and reached a maximum height of 0.6 stellar radii above its surface. The flare occurred around the south pole of Algol B, and energy must have been released continously throughout its life. We conclude that a specific extrastellar environment is not required for the presence of a flare, and that the processes at work are therefore similar to those on the Sun.Comment: Nature, Sept. 2 199

On the Contribution of Unresolved Galactic Stars to the Diffuse Soft X-ray Background

Using stellar luminosity functions derived from ROSAT data, the contributions of Galactic stars to the diffuse X-ray background are calculated for ROSAT PSPC energy bands. The model follows that of Guillout et al. (1996), but uses ROSAT rather than {\it Einstein} data to determine the intrinsic luminosity distributions. The model adequately predicts the numbers of stellar sources observed in deep ROSAT surveys. The contribution of unresolved stellar sources to the ROSAT All-Sky Survey at the Galactic poles is 6.85, 4.76, and 4.91 $\times$ counts s$^{-1}$ arcmin$^{-2}$ in bands R12 (1/4 keV), R45(3/4 keV), and R67(1.5 keV), respectively, which is equivalent to 4.66, 31.3 and 26.9 $\times10^{-14}$ ergs cm$^{-2}$ s$^{-1}$ deg$^{-2}$.Comment: 14 pages, 8 figures, accepted by Ap

The Sun as an X-ray Star: III. Flares

In previous works we have developed a method to convert solar X-ray data, collected with the Yohkoh/SXT, into templates of stellar coronal observations. Here we apply the method to several solar flares, for comparison with stellar X-ray flares. Eight flares, from weak (GOES class C5.8) to very intense ones (X9) are selected as representative of the flaring Sun. The emission measure distribution vs. temperature, EM(T), of the flaring regions is derived from Yohkoh/SXT observations in the rise, peak and decay of the flares. The EM(T) is rather peaked and centered around $T \approx 10^7$ K for most of the time. Typically, it grows during the rise phase of the flare, and then it decreases and shifts toward lower temperatures during the decay, more slowly if there is sustained heating. The most intense flare we studied shows emission measure even at very high temperature ($T \approx 10^8$ K). Time-resolved X-ray spectra both unfiltered and filtered through the instrumental responses of the non-solar instruments ASCA/SIS and ROSAT/PSPC are then derived. Synthesized ASCA/SIS and ROSAT/PSPC spectra are generally well fitted with single thermal components at temperatures close to that of the EM(T) maximum, albeit two thermal components are needed to fit some flare decays. ROSAT/PSPC spectra show that solar flares are in a two-orders of magnitude flux range ($10^6 - 10^8$ erg cm$^{-2}$ s$^{-1}$) and a narrow PSPC hardness ratio range, however higher than that of typical non-flaring solar-like stars.Comment: 32 pages, 8 figures, 3 table

Phospho-CRKL monitoring for the assessment of BCR-ABL activity in imatinib-resistant chronic myeloid leukemia or Ph+ acute lymphoblastic leukemia patients treated with nilotinib

n/

Coronal abundances from high-resolution X-ray data: The case of Algol

We discuss the determination of elemental abundances from high resolution X-ray data. We emphasize the need for an accurate determination of the underlying temperature structure and advocate the use of a line ratio method which allows us to utilize, first, the strongest lines observed in the X-ray spectra, and second, lines that span a rather wide temperature range. We point out the need to use continuous emission measure distributions and show via example that modeling in terms of individual temperature components yields errors of more than 50%. We stress the need to derive differential emission measure distributions based on physical assumptions and considerations. We apply our methods to the Chandra LETGS spectrum of Algol and show that nitrogen is considerably enhanced compared to cosmic abundances by a factor of 2 while carbon is depleted by at least a factor of 25. Iron, silicon, and magnesium, are all depleted compared to cosmic abundances, while the noble gas neon has the relatively highest abundance.Comment: 15 pages, 6 figures, A&A accepte