• Over 30% of patients with unexplained cytopenias who do not meet diagnostic criteria for MDS carry MDS-associated somatic mutations.• Clonal cytopenias of undetermined significance are more common than MDS and show comparable variant allele frequencies and blood counts.Establishing a diagnosis in patients suspected of having a myelodysplastic syndrome (MDS) can be challenging and could be informed by the identification of somatic mutations. We performed a prospective study to examine the frequency and types of mutations encountered in 144 patients with unexplained cytopenias. Based on bone marrow findings, 17% were diagnosed with MDS, 15% with idiopathic cytopenias of undetermined significance (ICUS) and some evidence of dysplasia, and 69% with ICUS and no dysplasia. Bone marrow DNA was sequenced for mutations in 22 frequently mutated myeloid malignancy genes. Somatic mutations were identified in 71% of MDS patients, 62% of patients with ICUS and some dysplasia, and 20% of ICUS patients and no dysplasia. In total, 35% of ICUS patients carried a somatic mutation or chromosomal abnormality indicative of clonal hematopoiesis. We validated these results in a cohort of 91 lower-risk MDS and 249 ICUS cases identified over a 6-month interval. Mutations were found in 79% of those with MDS, in 45% of those with ICUS with dysplasia, and in 17% of those with ICUS without dysplasia. The spectrum of mutated genes was similar with the exception of SF3B1 which was rarely mutated in patients without dysplasia. Variant allele fractions were comparable between clonal ICUS (CCUS) and MDS as were mean age and blood counts. We demonstrate that CCUS is a more frequent diagnosis than MDS in cytopenic patients. Clinical and mutational features are similar in these groups and may have diagnostic utility once outcomes in CCUS patients are better understood. (Blood. 2015;126(21):2355-2361 IntroductionMyelodysplastic syndromes (MDS) are clonal bone marrow disorders characterized by inefficient and dysmorphic hematopoietic differentiation, cytopenias of the peripheral blood, and increased risk of transformation to acute myeloid leukemia (AML).1 Establishing a diagnosis of MDS in a cytopenic patient is often challenging as the bone marrow must demonstrate dysplasia in 10% or more of a myeloid cell lineage or a blast proportion of 5% or greater.2 Quantification of these features can be subjective and prone to wide interobserver variation even among expert hematopathologists. 3,4 In cases that do not meet either bone marrow criteria, the presence of certain clonal karyotype abnormalities typical for MDS can serve as presumptive evidence of the diagnosis.2 Finally, other neoplasms and nonclonal causes of cytopenias must also be reasonably excluded. Many patients with otherwise unexplained cytopenias will fail to meet the diagnostic criteria for MDS and instead carry a designation of idiopathic cytopenias of undetermined significance (ICUS). 5-7The natural history of patients with ICUS is largely unknown and appears to be highly variable. Sma...

Conductive hydrogel scaffolds have important applications for electroactive tissue repairs. However, the development of conductive hydrogel scaffolds tends to incorporate nonbiodegradable conductive nanomaterials that will remain in the human body as foreign matters. Herein, a biodegradable conductive hybrid hydrogel is demonstrated based on the integration of black phosphorus (BP) nanosheets into the hydrogel matrix. To address the challenge of applying BP nanosheets in tissue engineering due to its intrinsic instability, a polydopamine (PDA) modification method is developed to improve the stability. Moreover, PDA modification also enhances interfacial bonding between pristine BP nanosheets and the hydrogel matrix. The incorporation of polydopamine-modified black phosphorous (BP@PDA) nanosheets into the gelatin methacryloyl (GelMA) hydrogels significantly enhances the electrical conductivity of the hydrogels and improves the cell migration of mesenchymal stem cells (MSCs) within the 3D scaffolds. On the basis of the gene expression and protein level assessments, the BP@PDAincorporated GelMA scaffold can significantly promote the differentiation of MSCs into neural-like cells under the synergistic electrical stimulation. This strategy of integrating biodegradable conductive BP nanomaterials within a biocompatible hydrogel provides a new insight into the design of biomaterials for broad applications in tissue engineering of electroactive tissues, such as neural, cardiac, and skeletal muscle tissues.

The myelodysplastic syndromes (MDSs) are collections of heterogeneous hematologic diseases characterized by refractory cytopenias as a result of ineffective hematopoiesis. Development of effective treatments has been impeded by limited insights into any unifying pathogenic pathways. We provide evidence that the p38 MAP kinase is constitutively activated or phosphorylated in MDS bone marrows. Such activation is uniformly IntroductionThe myelodysplastic syndromes (MDSs) comprise a spectrum of stem-cell malignancies characterized by cytologic dysplasia and ineffective hematopoiesis. [1][2][3] Although approximately one third of patients may experience progression to acute leukemia, refractory cytopenias are the principal cause of morbidity and mortality. MDS can be divided into low-and high-risk subtypes using the International Prognostic Scoring System (IPSS), based on features such as the number of hematopoietic deficits, the percentage of marrow blasts, and cytogenetic pattern. 4 Approximately two thirds of patients present with lower-risk disease (Low and Int-1 IPSS scores) characterized by increased rates of apoptosis in the progenitor and differentiated cell compartments in the marrow. [5][6][7][8] High intramedullary apoptosis leads to ineffective hematopoiesis and peripheral cytopenias. Higher grade or more advanced disease categories (Int-2 and High IPSS scores) are associated with a significant risk of leukemia transformation with a corresponding lower apoptotic index and higher percentage of marrow blasts.Cytokines play important roles in the regulation of normal hematopoiesis, and a balance between the actions of hematopoietic growth factors and myelosuppressive factors is required for optimal production of different hematopoietic-cell lineages. Excess production of inhibitory cytokines contributes in part to ineffective hematopoiesis in MDS. Tumor necrosis factor-␣ (TNF␣) has been implicated in the increased stem-cell apoptosis seen in MDS, 9,10 and high expression of TNF receptors and TNF mRNA have been reported in MDS bone marrows. [11][12][13][14] Transforming growth factor-␤ (TGF␤), interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), and interferon (IFN-␥ and -␣) are also myelosuppressive, and these proinflammatory cytokines have been found to be elevated in serum of patients with MDS in various studies and are hypothesized to play a role in suppressing hematopoiesis in this disease. 9,11,[15][16][17] Because multiple cytokines are involved in promoting abnormal hematopoietic development in MDS, targeting one single cytokine may not yield appreciable clinical benefit. In fact, anti-TNF therapeutic strategies (monoclonal antibodies and TNFR blockers) have only shown minimal efficacy. [18][19][20][21] Thus, it is imperative to identify common targetable pathways that regulate many different cytokines. Our previous studies have shown that myelosuppressive cytokines such as interferons (IFN-␣, -␤, and -␥), TGF␤, and TNF␣ can all activate the p38 mitogen activated protein kinase (MAPK) in ...

Purpose Myeloma-directed cellular immune responses after autologous stem cell transplantation (ASCT) may reduce relapse rates. We studied whether coinjecting the TLR-3 agonist and vaccine adjuvant Poly-ICLC with a MAGE-A3 peptide vaccine was safe and would elicit a high frequency of vaccine-directed immune responses when combined with vaccine-primed and costimulated autologous T cells. Experimental Design In a phase II clinical trial (NCT01245673), we evaluated the safety and activity of ex vivo expanded autologous T cells primed in vivo using a MAGE-A3 multipeptide vaccine (compound GL-0817) combined with Poly-ICLC (Hiltonol), granulocyte macrophage colony-stimulating factor (GM-CSF) ± montanide. Twenty-seven patients with active and/or high-risk myeloma received autografts followed by anti-CD3/anti-CD28–costimulated autologous T cells, accompanied by MAGE-A3 peptide immunizations before T-cell collection and five times after ASCT. Immune responses to the vaccine were evaluated by cytokine production (all patients), dextramer binding to CD8+ T cells, and ELISA performed serially after transplant. Results T-cell infusions were well tolerated, whereas vaccine injection site reactions occurred in >90% of patients. Two of nine patients who received montanide developed sterile abscesses; however, this did not occur in the 18 patients who did not receive montanide. Dextramer staining demonstrated MAGE-A3–specific CD8 T cells in 7 of 8 evaluable HLA-A2+ patients (88%), whereas vaccine-specific cytokine-producing T cells were generated in 19 of 25 patients (76%). Antibody responses developed in 7 of 9 patients (78%) who received montanide and only weakly in 2 of 18 patients (11%) who did not. The 2-year overall survival was 74% [95% confidence interval (CI), 54%–100%] and 2-year event-free survival was 56% (95% CI, 37%–85%). Conclusions A high frequency of vaccine-specific T-cell responses were generated after transplant by combining costimulated autologous T cells with a Poly-ICLC/GM-CSF–primed MAGE-A3 vaccine.

Advances in cardiovascular materials have brought us improved artificial vessels with larger diameters for reducing adverse responses that drive acute thrombosis and the associated complications. Nonetheless, the challenge is still considerable when applying these materials in small-diameter blood vessels. Here we report the biomimetic design of an acellular small-diameter vascular graft with specifically lamellar nanotopography on the luminal surface via a modified freeze-cast technique. The experimental findings verify that the well-designed nanolamellar structure is able to inhibit the adherence and activation of platelets, induce oriented growth of endothelial cells, and eventually remodel a neovessel to maintain long-term patency in vivo. Furthermore, the results of numerical simulations in physically mimetic conditions reveal that the regularly lamellar nanopattern can manipulate blood flow to reduce the flow disturbance compared with random topography. Our current work not only creates a freeze-cast small-diameter vascular graft that employs topographic architecture to direct the vascular cell fates for revasculature but also rekindles confidence in biophysical cues for modulating in situ tissue regeneration.

A melamine-modified fibroin was synthesized and fabricated into electrospun nanofiber films with broad-spectrum antimicrobial activity, sustained water retention, and fast reepithelialization and revascularization.

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The epidermal growth factor receptor (EGFR) family belongs to type I receptor tyrosine kinases. Overexpression or mutation of EGFR/ErbB1 gene has been detected in a large number of human solid tumours. According to some previous report, this gene is not expressed in hematological malignancies. However, two recent clinical case reports showed that erlotinib caused complete remission of acute myeloid leukaemia (AML)-M1 in patients who had both AML-M1 and non-small-cell lung cancer. These results are supported by preclinical studies in which EGFR tyrosine kinase inhibitors have anti-proliferative effects on AML. These findings prompted us to determine whether EGFR is expressed in human AML, through a large-scale screening of both leukaemic cell lines and clinical samples. Our results show that EGFR is expressed by about 33% of human AML (containing M1 to M7 subtypes) and by some human leukaemia cell lines (K562, MEG-01, CEM and SKO-007). Its expression is not limited to certain AML types but has been detected in many leukaemic cells. In addition, EGFR expression was intimately associated with the poor clinical outcomes. Finally, we find that only EGFR-positive leukaemic cells respond to antibody-dependent cellular cytotoxicity of cetuximab, the monoclonal antibodies against EGFR.