The Bcl-2 inhibitor venetoclax inhibits Nrf2 antioxidant pathway activation induced by hypomethylating agents in AML.

Induction of reactive oxygen species (ROS), an important process for the cytotoxicity of various acute myeloid leukemia (AML) therapies including hypomethylating agents (HMAs), concurrently activates the NF-E2-related factor 2 (Nrf2) antioxidant response pathway which in turn results in induction of antioxidant enzymes that neutralize ROS. In this study, we demonstrated that Nrf2 inhibition is an additional mechanism responsible for the marked antileukemic activity in AML seen with the combination of HMAs and venetoclax (ABT-199). HMA and venetoclax combined treatment augmented mitochondrial ROS induction and apoptosis compared with treatment HMA alone. Treatment of AML cell lines as well as primary AML cells with venetoclax disrupted HMA decitabine-increased nuclear translocation of Nrf2 and induction of downstream antioxidant enzymes including heme oxygenase-1 and NADP-quinone oxidoreductase-1. Venetoclax treatment also leads to dissociation of B-cell lymphoma 2 from the Nrf2/Keap-1 complex and targets Nrf2 to ubiquitination and proteasomal degradation. Thus, our results here demonstrated an undiscovered mechanism underlying synergistic effect of decitabine and venetoclax in AML cells, elucidating for impressive results in antileukemic activity against AML in preclinical and early clinical studies by combined treatment of these drugs.

8-chloro-adenosine activity in FLT3-ITD acute myeloid leukemia.

Nucleoside analogs represent the backbone of several distinct chemotherapy regimens for acute myeloid leukemia (AML) and combination with tyrosine kinase inhibitors has improved survival of AML patients, including those harboring the poor-risk FLT3-ITD mutation. Although these compounds are effective in killing proliferating blasts, they lack activity against quiescent leukemia stem cells (LSCs), which contributes to initial treatment refractoriness or subsequent disease relapse. The reagent 8-chloro-adenosine (8-Cl-Ado) is a ribose-containing, RNA-directed nucleoside analog that is incorporated into newly transcribed RNA rather than in DNA, causing inhibition of RNA transcription. In this report, we demonstrate antileukemic activities of 8-Cl-Ado in vitro and in vivo and provide mechanistic insight into the mode of action of 8-Cl-Ado in AML. 8-Cl-Ado markedly induced apoptosis in LSC, with negligible effects on normal stem cells. 8-Cl-Ado was particularly effective against AML cell lines and primary AML blast cells harboring the FLT3-ITD mutation. FLT3-ITD is associated with high expression of miR-155. Furthermore, we demonstrate that 8-Cl-Ado inhibits miR-155 expression levels accompanied by induction of DNA-damage and suppression of cell proliferation, through regulation of miR-155/ErbB3 binding protein 1(Ebp1)/p53/PCNA signaling. Finally, we determined that combined treatment of NSG mice engrafted with FLT3-ITD + MV4-11 AML cells with 8-Cl-Ado and the FLT3 inhibitor AC220 (quizartinib) synergistically enhanced survival, compared with that of mice treated with the individual drugs, suggesting a potentially effective approach for FLT3-ITD AML patients.

Exosome-mediated microenvironment dysregulation in leukemia.

The hematopoietic stem cell (HSC) niche is composed of a complex set of stromal support cells that maintain HSCs and promote normal hematopoiesis. We now know that molecular changes within the hematopoietic niche contribute to leukemia development. Leukemia cells often reorganize the hematopoietic niche to promote and support their own survival and growth. Here we will summarize recent works that decipher the normal hematopoietic niche cellular components and describe how the leukemia-transformed niche contributes to hematological malignances. Finally, we will discuss recent publications that highlight a possible role for exosomes in the leukemia-induced niche reorganization. This article is part of a Special Issue entitled: Tumor Microenvironment Regulation of Cancer Cell Survival, Metastasis, Inflammation, and Immune Surveillance edited by Peter Ruvolo and Gregg L. Semenza.

Altered lymphopoiesis and immunodeficiency in miR-142 null mice.

MicroRNAs (miRNAs) are a class of powerful posttranscriptional regulators implicated in the control of diverse biological processes, including regulation of hematopoiesis and the immune response. To define the biological functions of miR-142, which is preferentially and abundantly expressed in immune cells, we created a mouse line with a targeted deletion of this gene. Our analysis of miR-142(-/-) mice revealed a critical role for this miRNA in the development and homeostasis of lymphocytes. Marginal zone B cells expand in the knockout spleen, whereas the number of T and B1 B cells in the periphery is reduced. Abnormal development of hematopoietic lineages in miR-142(-/-) animals is accompanied by a profound immunodeficiency, manifested by hypoimmunoglobulinemia and failure to mount a productive immune response to soluble antigens and virus. miR-142(-/-) B cells express elevated levels of B-cell-activating factor (BAFF) receptor (BAFF-R) and as a result proliferate more robustly in response to BAFF stimulation. Lowering the BAFF-R gene dose in miR-142(-/-) mice rescues the B-cell expansion defect, suggesting that BAFF-R is a bona fide miR-142 target through which it controls B-cell homeostasis. Collectively, our results uncover miR-142 as an essential regulator of lymphopoiesis, and suggest that lesions in this miRNA gene may lead to primary immunodeficiency.

Biobased, cellulose long filament-reinforced vanillin-derived epoxy composite for high-performance and flame-retardant applications.

The development of high-strength and intrinsic flame-retardant natural fiber-reinforced green composite (NFRGC) is a landmark for high-performance structural applications. This paper reports a biobased, high-performance, flame-retardant composite material based on diverse bio-resources. Tough and strong cellulose long filaments (CLFs) are combined with vanillin-derived epoxy (VDE) resin to achieve high strength and flame-retardant NFRGC. The green composite was fabricated using a simple hand lay-up and compression molding technique. The green composite showed a noteworthy increment of 100.9 % flexural strength and 346 % flexural modulus compared to the neat VDE resin. Interestingly, despite the highly flammable nature of CLF, the green composite passes a V-0 rating under the UL-94 test, indicating excellent flame-retardant characteristics. Additionally, the green composite demonstrated outstanding hydrophobicity with a water contact angle of 104.2° and good chemical stability in various acidic and organic solvents. The green composite's excellent mechanical and physical properties show its potential for high-strength and flame-retardant structural applications.

Advanced hydrostable, recyclable and degradable cellulose hybrid films as renewable alternatives to synthetic plastics.

Strong, tough and sustainable materials are in high demand in various engineering applications. We demonstrate a potential sustainable hybrid film made from natural cellulose and a biobased slurry. Through a simple and scalable approach, cellulose can be processed into an advanced material with over 2.8 and 9.2-fold increase in dry strength and toughness after curing and a 728-fold increase in wet strength, respectively. In addition, these hybrid composite films display an outstanding antioxidant activity surpassing 90 %, along with excellent ultraviolet radiation shielding and thermal insulation properties. Further, the hybrid films can be fabricated by integrating all-natural materials and still guarantee their unique functionality. We also demonstrate the feasibility of a circular bioeconomy by recycling the hybrid film using a green, deep eutectic solvent to fabricate a recycled hybrid film that displays excellent mechanical and optical properties. When recycling is unsuitable or economical, the hybrid film can naturally degrade in the soil under 6 months. These encouraging findings suggest the promise of cellulose hybrid films as a renewable, low-cost, tough, and strong material with the potential to replace nonrenewable synthetic plastics and products.

Environment-friendly, high-performance cellulose nanofiber-vanillin epoxy nanocomposite with excellent mechanical, thermal insulation and UV shielding properties.

With the increased demand for biobased epoxy thermosets as an alternative to petroleum-based materials in various fields, developing environment-friendly and high-performance natural fiber-biobased epoxy nanocomposites is crucial for industrial applications. Herein, an environment-friendly nanocomposite is reported by introducing cellulose nanofiber (CNF) in situ interaction with lignin-derived vanillin epoxy (VE) monomer and 4, 4´-diaminodiphenyl methane (DDM) hardener that serves as a multifunctional platform. The CNF-VE nanocomposite is fabricated by simply dispersing the CNF suspension to the VE and DDM hardener solution through the in-situ reaction, and its mechanical properties and thermal insulation behavior, wettability, chemical resistance, and optical properties are evaluated with the CNF weight percent variation. The well-dispersed CNF-VE nanocomposite exhibited high tensile strength (∼127.78 ± 3.99 MPa) and strain-at-break (∼16.49 ± 0.61 %), haziness (∼50 %) and UV-shielding properties. The in situ loading of CNF forms covalent crosslinking with the VE and favors improving the mechanical properties along with the homogeneous dispersion of CNF. The CNF-VE nanocomposite also shows lower thermal conductivity (0.26 Wm-1K-1) than glass. The environment-friendly and high-performance nanocomposite provides multiple platforms and can be used for building materials.

Strong, highly porous and sustainable nanocellulose foam made using bioderived hyperbranched crosslinker for thermal insulation and sound absorption.

This paper reports an environment-friendly biobased foam made with cellulose nanofiber (CNF) and a biobased hyperbranched crosslinker, glycerol succinic anhydride (GSA). As a biobased hyperbranched crosslinker, carboxyl-terminated GSA is synthesized through a straightforward esterification process involving glycerol and succinic anhydride. The GSA-crosslinked CNF (GSA/CNF) foam is prepared using a facile, sustainable, cost-effective, and efficient solvent-exchange method. The resulting foam exhibits notable characteristics, including improved dimensional stability, remarkably low density (13.41 mg/cm3) with high porosity (>99 %), and exceptional compressive strength (494 kPa) and modulus (452 kPa). Further, the foam offers outstanding sound absorption capabilities with a coefficient of 0.986 at 2 kHz and remarkably low thermal conductivity (30.18 mW/mK), significantly lower than commonly used and reported porous materials, indicating its potential as an efficient, environmentally friendly sound absorption and thermal insulation material.

Single-, Dual-, and Multi-Stimuli-Responsive Nanogels for Biomedical Applications.

In recent years, stimuli-responsive nanogels that can undergo suitable transitions under endogenous (e.g., pH, enzymes and reduction) or exogenous stimuli (e.g., temperature, light, and magnetic fields) for on-demand drug delivery, have received significant interest in biomedical fields, including drug delivery, tissue engineering, wound healing, and gene therapy due to their unique environment-sensitive properties. Furthermore, these nanogels have become very popular due to some of their special properties such as good hydrophilicity, high drug loading efficiency, flexibility, and excellent biocompatibility and biodegradability. In this article, the authors discuss current developments in the synthesis, properties, and biomedical applications of stimulus-responsive nanogels. In addition, the opportunities and challenges of nanogels for biomedical applications are also briefly predicted.

TP-0184 inhibits FLT3/ACVR1 to overcome FLT3 inhibitor resistance and hinder AML growth synergistically with venetoclax.

We identified activin A receptor type I (ACVR1), a member of the TGF-ß superfamily, as a factor favoring acute myeloid leukemia (AML) growth and a new potential therapeutic target. ACVR1 is overexpressed in FLT3-mutated AML and inhibition of ACVR1 expression sensitized AML cells to FLT3 inhibitors. We developed a novel ACVR1 inhibitor, TP-0184, which selectively caused growth arrest in FLT3-mutated AML cell lines. Molecular docking and in vitro kinase assays revealed that TP-0184 binds to both ACVR1 and FLT3 with high affinity and inhibits FLT3/ACVR1 downstream signaling. Treatment with TP-0184 or in combination with BCL2 inhibitor, venetoclax dramatically inhibited leukemia growth in FLT3-mutated AML cell lines and patient-derived xenograft models in a dose-dependent manner. These findings suggest that ACVR1 is a novel biomarker and plays a role in AML resistance to FLT3 inhibitors and that FLT3/ACVR1 dual inhibitor TP-0184 is a novel potential therapeutic tool for AML with FLT3 mutations.

Increased expression of CD70 in relapsed acute myeloid leukemia after hypomethylating agents.

Acute myeloid leukemia (AML) is the most common acute leukemia in adults. While induction chemotherapy leads to remission in most patients, a significant number will experience relapse. Therefore, there is a need for novel therapies that can improve remission rates in patients with relapsed and refractory AML. CD70 is the natural ligand for CD27 (a member of the TNF superfamily) and appears to be a promising therapeutic target. Consequently, there is considerable interest in developing chimeric antigen receptor (CAR) T-cell therapy products that can specifically target CD70 in various neoplasms, including AML. In this study, we employed routine diagnostic techniques, such as immunohistochemistry and flow cytometry, to investigate the expression of CD70 in bone marrow samples from treatment-naïve and relapsed AML patients after hypomethylating agents (HMA). Also, we evaluated the impact of HMA on CD70 expression and examined CD70 expression in various leukemic cell subsets and normal hematopoietic progenitors.

CD28 costimulation augments CAR signaling in NK cells via the LCK/CD3Z/ZAP70 signaling axis.

Multiple factors in the design of a chimeric antigen receptor (CAR) influence CAR T-cell activity, with costimulatory signals being a key component. Yet, the impact of costimulatory domains on the downstream signaling and subsequent functionality of CAR-engineered natural killer (NK) cells remains largely unexplored. Here, we evaluated the impact of various costimulatory domains on CAR-NK cell activity, using a CD70-targeting CAR. We found that CD28, a costimulatory molecule not inherently present in mature NK cells, significantly enhanced the antitumor efficacy and long-term cytotoxicity of CAR-NK cells both in vitro and in multiple xenograft models of hematologic and solid tumors. Mechanistically, we showed that CD28 linked to CD3Z creates a platform that recruits critical kinases, such as LCK and ZAP70, initiating a signaling cascade that enhances CAR-NK cell function. Our study provides insights into how CD28 costimulation enhances CAR-NK cell function and supports its incorporation in NK-based CARs for cancer immunotherapy.

Mechanical and Dynamic Mechanical Behavior of the Lignocellulosic Pine Needle Fiber-Reinforced SEBS Composites.

Aiming to generate wealth from waste and due to their significant fire threats to forests and their rich cellulose content, lignocellulosic pine needle fibers (PNFs) are utilized in this study as a reinforcement of the thermoplastic elastomer styrene ethylene butylene styrene (SEBS) matrix to create environmentally friendly and economical PNF/SEBS composites using a maleic anhydride-grafted SEBS compatibilizer. The chemical interaction in the composites studied by FTIR shows that strong ester bonds are formed between reinforcing PNF, the compatibilizer, and the SEBS polymer, leading to strong interfacial adhesion between the PNF and SEBS in the composites. This strong adhesion in the composite exhibits higher mechanical properties than the matrix polymer indicating a 1150 % higher modulus and a 50 % higher strength relative to the matrix. Further, the SEM pictures of the tensile-fractured samples of the composites validate this strong interface. Finally, the prepared composites show better dynamic mechanical behavior indicating higher storage and loss moduli and Tg than the matrix polymer suggesting their potential for engineering applications.

Development and Evaluation of Cellulose/Graphene-Oxide Based Composite for Removing Phenol from Aqueous Solutions.

In this study, a graphene oxide/cellulose composite (GO-cellulose) was prepared usingcellulose and graphene oxide (GO) through ultrasonication, followed by the freeze-dried method. The Brunauer-Emmett-Teller (BET) specific surface area of GO-cellulose (~6.042 m2/g) was higher compared to cellulose (1.023 m2/g).The UV-Visible spectraindicated that the prepared GO-cellulose composite removedphenol efficiently from aqueous solutions with high adsorption power. The effectiveness of the composite for phenol adsorption was examinedunder diverse conditions.The results reveal that the composite optimally improved the adsorption at pH 7 with a dose of 0.125 g/30 L in about 40 min. The adsorption process showed that in optimum conditions, 86 ± 2% of phenol was removed in 40 min with an adsorption capacity of 6.192 mg g-1. The adsorption behavior was well fitted to the pseudo-second-order kinetic model and the Langmuir isotherms at all temperatures.The present study suggests that synthesized GO-cellulose is useful inthe removal of phenol pollutants from aqueous solutions.

Fixation of Fresh Femoral Neck Fractures Using Fibular Strut Graft Along with Cannulated Screws.

BACKGROUND: Intracapsular femoral neck fractures can result from trivial trauma in the elderly or high-energy trauma in younger age-groups. In younger patients aged <60 years, the femoral head should be conserved to avoid the long-term complications of replacement arthroplasty. The options of osteosynthesis include closed/ open reduction and internal fixation with/without bone grafting. Internal fixation alone does not provide rigid fixation, owing to cavities in the posterior part of the femoral head and neck. Fibular grafts augment union and provide strength to the posterior cortex during reconstruction of the femoral neck. We evaluated the use of fibular grafting for fresh femoral neck fractures with posterior comminution. MATERIAL AND METHODS: Between November 2019 and March 2022, 20 women and 12 men aged 20 to 60 years underwent osteosynthesis and fibular strut grafting supplemented with 7.0-mm cannulated hip screws for Garden grades III (n=19) and IV (n=13) femoral neck fractures. Clinical and radiological outcomes were evaluated. RESULTS: Patients were followed up for a period of 15 months. According to the Harris hip score, outcome was good to excellent in 23, fair in 7, and poor in 2. 30 of the 32 patients achieved bone union after a mean of 4.5 (range 3.5-5.5) months. In 2 patients, the bone was united with a mean of 10º of varus collapse. Two patients had non-union. Other complications included screw migration in the joint space (n=1) and screw pullout (n=2). No patient had avascular necrosis of the femoral head. CONCLUSION: Fixation with cancellous screws and fibular strut grafts for femoral neck fractures is cost-effective and technically less demanding, and associated with good outcomes.

Early Results of Internal Fixation with Femoral Neck System in Young Patients with Femoral Neck Fracture.

BACKGROUND: Femoral neck fractures are intracapsular hip fractures. There are several surgical implants that have been used to treat femoral neck fractures. Depuy Synthes Products and the lower extremity expert group have developed an innovative femoral neck system (FNS) for fixing femoral neck fractures. With minimally invasive procedures, FNS can provide angular stability. MATERIALS AND METHODS: A non-randomized single centre prospective study was conducted in 30 patients of less than 60 years of age at Maharishi Markandeshwar Institute of Medical Sciences and Research between August 2020 to May 2022. All patients underwent internal fixation with FNS within 48 hours of presentation. RESULTS: Although blood loss and operative time in our operated group was more than that in conventional fixation by cannulated screws, our group had better VAS scores, better Harris scores and lower complication rates. CONCLUSIONS: 1. The Femoral Neck System resulted in better biomechanical properties and good early results in femoral neck fractures. FNS gives both angular stability and rotational stability. The last follow-up in our study showed Harris score to be significantly higher, and the incidence of complications were lower. 2. The combination of FNS bolts with anti-rotation screws avoids the "Z" effect and improves the overall stability and anti-rotation effect. 3. In addition, the novel sliding compression mechanism of FNS allows the fracture ends to come in close contact with each other, benefitting fracture healing.

Effect of bio-based derived epoxy resin on interfacial adhesion of cellulose film and applicability towards natural jute fiber-reinforced composites.

This paper reports a bio-based vanillin-derived epoxy (VDE) resin for bio-based natural fiber-reinforced composites. VDE monomer was synthesized, and curing agents, namely, 4,4´-diaminodiphenyl methane (DDM) and isophorone diamine, were used. The prepared VDE resins with various curing parameters were characterized using FTIR, NMR, tensile test, bending test and water contact angle. Further, the interfacial adhesion feasibility of VDE resins on cellulose film was studied through the single-lap shear joint examination and compared with a commercial epoxy, DGEBA. The VDE-DDM resin exhibited excellent interfacial adhesion with cellulose than VDE-IPDA and DGEBA-DDM resins. The cured VDE-DDM thermoset showed a tensile strength of 86.0 ± 6.5 MPa, thermal stability of 241.0 °C at Td5%, and an elastic modulus of 2.9 ± 0.3 GPa, which is better than the commercial epoxy resin. Besides, the developed VDE-DDM resin was used to fabricate treated-jute fiber (TJF)-reinforced composites. The bio-based VDE-DDM/TJF composite's flexural strength was higher than the commercial epoxy resin composite, DGEBA-DDM/TJF. Furthermore, the phosphorus moiety of the VDE-DDM resin endows flame retardancy to the VDE-DDM/TJF composite during combustion. Overall, the appealing properties of bio-based VDE-DDM/TJF composite render environment-friendly and high-performance structural applications.

Engineered cord blood megakaryocytes evade killing by allogeneic T-cells for refractory thrombocytopenia.

The current global platelet supply is often insufficient to meet all the transfusion needs of patients, in particular for those with alloimmune thrombocytopenia. To address this issue, we have developed a strategy employing a combination of approaches to achieve more efficient production of functional megakaryocytes (MKs) and platelets collected from cord blood (CB)-derived CD34+ hematopoietic cells. This strategy is based on ex-vivo expansion and differentiation of MKs in the presence of bone marrow niche-mimicking mesenchymal stem cells (MSCs), together with two other key components: (1) To enhance MK polyploidization, we used the potent pharmacological Rho-associated coiled-coil kinase (ROCK) inhibitor, KD045, resulting in liberation of increased numbers of functional platelets both in-vitro and in-vivo; (2) To evade HLA class I T-cell-driven killing of these expanded MKs, we employed CRISPR-Cas9-mediated ß-2 microglobulin (ß2M) gene knockout (KO). We found that coculturing with MSCs and MK-lineage-specific cytokines significantly increased MK expansion. This was further increased by ROCK inhibition, which induced MK polyploidization and platelet production. Additionally, ex-vivo treatment of MKs with KD045 resulted in significantly higher levels of engraftment and donor chimerism in a mouse model of thrombocytopenia. Finally, ß2M KO allowed MKs to evade killing by allogeneic T-cells. Overall, our approaches offer a novel, readily translatable roadmap for producing adult donor-independent platelet products for a variety of clinical indications.

Redox-sensitive nanoparticles based on xylan-lipoic acid conjugate for tumor targeted drug delivery of niclosamide in cancer therapy.

In this study, novel redox-sensitive nanoparticles based on xylan-lipoic acid (Xyl-LA) conjugate were developed for tumor targeted delivery of niclosamide (Nic) in cancer therapy. The niclosamide loaded xylan-lipoic acid conjugate nanoparticles (Xyl-LA/Nic NPs) showed redox responsive behaviour in presence of reductive glutathione (GSH), which indicate their suitability for intracellular drug release. The obtained Xyl-LA/Nic NPs exhibited uniform particle size (196 ± 1.64 nm), high loading capacity (~28.6 wt %) and excellent blood compatibility. The anticancer activity of the Niclosamide and the Xyl-LA/Nic NPs against the colon carcinoma cell lines (HCT-15, Colo-320) were evaluated by MTT assay and the overall results indicate that the Xyl-LA/Nic NPs significantly enhanced the therapeutic efficiency of niclosamide in cancer therapy.