Search

Supplementary Data from Entinostat Decreases Immune Suppression to Promote Antitumor Responses in a HER2+ Breast Tumor Microenvironment

Metastatic breast cancer remains one of the leading causes of global cancer incidence in women, despite the benefit of immune checkpoint inhibitors (ICIs) in managing various cancers and improving patient quality of life. Metastatic breast cancer is characterized by extensive infiltration of the tumor microenvironment (TME) with immunosuppressive cells, such as myeloid derived suppressor cells (MDSCs), that inhibit anti-tumoral immune cells and prevent effective activation of the adaptive immune system by ICIs. Previously, our group has shown in the breast TME that epigenetic reprogramming of MDSCs by entinostat, a histone deacetylase inhibitor, decreases MDSC immunosuppressive function and enhances response to ICIs, in part mediated by altered signaling within the Signal transducer and activator of transcription 3 (STAT3) and Nuclear factor kappa B (NFκB) axis. Next, we sought to examine the effects of entinostat on MDSC reprogramming in a distant metastatic TME. Using the syngeneic NT2.5LM NeuN mouse model of metastatic breast cancer, we established spontaneous lung metastases and treated with either vehicle or entinostat (5 mg/kg by oral gavage, 5x/week) for 3 weeks. Single cell RNA sequencing (scRNAseq) of macro-dissected lung metastases revealed a large MDSC population, and unsupervised pathways analysis of the MDSC cluster showed differential regulation of the IL6-JAK-STAT3 and TNFα-signaling-via-NFκB pathways upon entinostat treatment. In line with reports of crosstalk among STAT3, NFκB, and AP-1 pathways regulating inflammation in breast cancer, we also found through scRNAseq differential expression of AP-1 subunits JunB and FOSL1 upon entinostat treatment. At the protein level, western blot analysis of isolated intratumoral MDSCs from lung metastases revealed decreased STAT3 phosphorylation upon entinostat treatment. Using J774M cells, an MDSC-like cell line, we found decreased JunB phosphorylation and decreased FOSL1 protein upon entinostat treatment. Furthermore, preliminary evaluation of imaging mass cytometry (IMC) from tumor biopsies in selected patients with metastatic breast cancer treated with entinostat during the clinical trial NCI-9844 confirmed decreased STAT3 phosphorylation in MDSCs. Taken together, we provide new evidence in the metastatic lung TME that implicates a STAT3-NFκB-AP-1 mediated mechanism leading to decreased MDSC suppression. Evaluating this mechanism in MDSCs taken directly from treated lung metastases represents the most biologically relevant mechanistic study to date, and preliminary translation of findings in patients suggests that planned studies will lead to identification of the mechanism driving response. Citation Format: Aaron G. Baugh, Edgar Gonzalez, Valerie H. Narumi, Sofi Castanon, Jesse Kreger, James Leatherman, Won Jin Ho, Ashley Cimino-Mathews, Vered Stearns, Roisin M. Connolly, Elizabeth M. Jaffee, Adam L. MacLean, Evanthia T. Roussos Torres. Entinostat's modulation of myeloid derived suppressor cells through the STAT3-NFκB-AP-1 axis decreases suppressive signaling. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5167.

Therapeutic combinations to alter immunosuppressive, solid tumor microenvironments (TME), such as in breast cancer, are essential to improve responses to immune checkpoint inhibitors (ICI). Entinostat, an oral histone deacetylase inhibitor, has been shown to improve responses to ICIs in various tumor models with immunosuppressive TMEs. The precise and comprehensive alterations to the TME induced by entinostat remain unknown. Here, we employed single-cell RNA sequencing on HER2-overexpressing breast tumors from mice treated with entinostat and ICIs to fully characterize changes across multiple cell types within the TME. This analysis demonstrates that treatment with entinostat induced a shift from a protumor to an antitumor TME signature, characterized predominantly by changes in myeloid cells. We confirmed myeloid-derived suppressor cells (MDSC) within entinostat-treated tumors associated with a less suppressive granulocytic (G)-MDSC phenotype and exhibited altered suppressive signaling that involved the NFκB and STAT3 pathways. In addition to MDSCs, tumor-associated macrophages were epigenetically reprogrammed from a protumor M2-like phenotype toward an antitumor M1-like phenotype, which may be contributing to a more sensitized TME. Overall, our in-depth analysis suggests that entinostat-induced changes on multiple myeloid cell types reduce immunosuppression and increase antitumor responses, which, in turn, improve sensitivity to ICIs. Sensitization of the TME by entinostat could ultimately broaden the population of patients with breast cancer who could benefit from ICIs.

The Valsartan Amlodipine Randomized Trial (VART) was performed to compare the beneficial effects of valsartan and amlodipine on cardiovascular events in Japanese hypertensive patients. In this subanalysis of the VART, we assessed the relationship between home blood pressure (HBP) levels and cardiovascular events in the enrolled patients. We enrolled 1021 patients with mild-to-moderate hypertension in the VART. The participants were allocated randomly to either the valsartan group or the amlodipine group. The primary end point was a composite of all-cause death, sudden death, cerebrovascular events, cardiac events, vascular events and renal events. A total of 621 patients (valsartan group: 305 and amlodipine group: 316) completed the measurements of HBP (morning and evening) throughout the trial. Both the agents evenly and significantly lowered morning HBP and evening HBP throughout the trial. There was no significant difference in the primary end point between the two groups. However, we observed significant decreases in the left ventricular mass index and urinary albumin to creatinine ratio in the valsartan group but not in the amlodipine group. There were no significant differences in HBP levels and the main outcome of the cardiovascular events between the valsartan and amlodipine groups. However, in the valsartan group, significant improvements in left ventricular hypertrophy and microalbuminuria were observed.

The oxidation wave of CO preadsorbed at 50 mV on Pt(1 1 1), (1 0 0) and (1 1 0) electrodes in phosphate buffer solution of pH 3 was observed as a function of the sweep rate. The sweep rate dependence of the peak current and peak potential, as well as the form of the wave, were examined on the basis of the Gilman mechanism that the electron transfer from a complex consisting of CO and oxygen containing species is the rate-determining step. An electron transfer step from CO itself was excluded. The peak current and peak potential analyses and the wave simulation gave the same value for Δf, the change in the interaction energy during the formation of the activated complex from the reactants. Δf was sweep-rate and surface-structure dependent. The nature of Δf was discussed.

The infrared astronomy satellite AKARI has made all‐sky surveys at six wavelength bands (9, 18 μm with the Infrared Camera (IRC), 65, 90, 140, and 160 μm with the Far‐Infrared Surveyor (FIS)). The first version of the FIS Bright Source Catalogue (β−1) has been provided to the AKARI science team for initial astronomical analyses. The catalogue will be made public in Autumn 2009 after further revisions. The IRC point source catalogue is in parallel preparation.

The eikonal partial wave theory for the electron impact excitation of positive ions is formulated and then demonstrated in the 1s to 2s, 2p excitations of hydrogen-like ions.

Holographic versatile disc (HVDtrade) evaluation system based on Collineartrade Technology, which is used to evaluate a rotating HVDtrade, is proposed. This system can easily be used to evaluate the specification of media or HVDtrade drive system

A separation optical setup for holographic versatile disc (HVDtrade) drive system, using Collineartrade Technology, is proposed. With this unique designed system configuration the pick-up can read or write holograms from any position on the rotating HVDtrade media

High density data recording of holographic versatile disc was demonstrated using collinear holographic technology in static mode. The optical noise reduction and improvement of the photopolymer characteristics is the key to the success.

Technologies that enable traceability for fishery products are increasing their demands. Recently proposed technologies are mainly based on disposal RF(IC) tags which are able to record information directly onto them. However, the current systems based on RF tags have problems of expensive price of tags, and weakness of reading information if applied onto surface of products containing much water, which prevents to construct practically feasible systems using the RF tags. To provide a traceability system that uses much inexpensive media and that assures as high security as the RF tags, we propose a system based on a combination of printed 2D codes and internet connection, with security control similar to on-line electronic transactions. The proposed system identifies a fishery product by giving it a unique serial ID, which is issued by a database server, and printed in 2D code onto a paper or a plastic plate, which is directly put on the product. All the trace information sent from client (producer, transporters, and retailer) via internet is associated to the ID and stored to the server. Since 2D code is able to be read by such as mobile phones with built-in camera, a consumer is able to get history of the product with a single scanning operation. For the weakness of printed codes against duplication by copying, we propose a method to identify its validity by digital encryption, along with identification by weight information. The system is assured its usability by a series of experiments conducted for the distribution of cultured flounder in Hakodate, Japan

Radicicol, a macrocyclic antifungal antibiotic, has been shown to bind to the heat shock protein 90 (Hsp90) chaperone, interfering with its function. Hsp90 family chaperones have been shown to associate with several signaling molecules and play an essential role in signal transduction, which is important for tumor cell growth. Because radicicol lacks antitumor activity in vivo in experimental animal models, we examined the antitumor activity of a novel radicicol oxime derivative, radicicol 6-oxime (KF25706), on human tumor cell growth both in vitro and in vivo. KF25706 showed potent antiproliferative activities against various human tumor cell lines in vitro and inhibited v-src- and K-ras-activated signaling as well as radicicol. In addition, Hsp90 family chaperone-associated proteins, such as p185erbB2, Raf-1, cyclin-dependent kinase 4, and mutant p53, were depleted by KF25706 at a dose comparable to that required for antiproliferative activity. KF25706 was also shown to compete with geldanamycin for binding to Hsp90. KF29163, which is an inactive derivative of radicicol, was less potent both in p185erbB2 depletion and Hsp90 binding. More importantly, KF25706 showed significant growth-inhibitory activity against human breast carcinoma MX-1 cells transplanted into nude mice at a dose of 100 mg/kg twice daily for five consecutive i.v. injections. KF25706 was also shown to possess antitumor activity against human breast carcinoma MCF-7, colon carcinoma DLD-1, and vulval carcinoma A431 cell lines in vivo in an animal model. Finally, we confirmed the depletion of Hsp90-associated signaling molecules (Raf-1 and cyclin-dependent kinase 4) with ex vivo Western blotting analysis using MX-1 xenografts. In agreement with in vivo antitumor activity, KF25706 depleted Hsp90-associated molecules in vivo, whereas KF29163 and radicicol did not show this activity in vivo. Taken together, these results suggest that antitumor activity of KF25706 may be mediated, at least in part, by binding to Hsp90 family proteins and destabilization of Hsp90-associated signaling molecules.

Engraftment failure following allogeneic bone marrow transplantation (BMT) is rare in patients with acute leukemia, after frequent conditioning with marrow-lethal chemoradiotherapy. We evaluated the efficacy of a preparatory regimen consisting of fractionated total body irradiation (TBI) (12 Gy in six fractions) and high dose etoposide (60 mg/kg) administered as an 8-h infusion for allogeneic BMT in 16 consecutive patients with acute leukemia. Although 14 patients showed complete and sustained engraftment, the remaining two patients rejected bone marrow grafts from HLA-identical sibling donors and showed subsequent recovery of host-derived hematopoiesis. Despite the limited number of patients, this observation suggests that the immunosuppressive potential of etoposide may be inferior to that of cyclophosphamide (CY) and that etoposide as an alternative to CY as an antileukemic and immunosuppressive agent in allogeneic BMT may increase the risk of graft rejection.

We evaluated a combination chemotherapy with cytarabine ocfosfate (SPAC), low dose etoposide and G-CSF for the treatment of high-risk MDS (RAEB, RAEBt). Seven patients with high-risk MDS were treated with a daily combination, 200 mg/day SPAC p.o., 50 mg/day etoposide p.o. and 75 micrograms/day G-CSF s.c. One patient achieved complete response, 2 achieved good response and one patient minor response. Although all of the patients developed severe marrow hypoplasia after chemotherapy, the nonhematologic adverse effects were mild enough to be tolerated. This combination chemotherapy should be useful in the clinical management of patients with high-risk MDS.

We describe the first case of T-cell prolymphocytic leukaemia (T-PLL) in which the peripheral blood cells contained a human T-lymphotropic virus (HTLV) related tax sequence. Serum screening tests for anti-HTLV-I/II antibodies were negative. Polymerase chain reaction disclosed the presence of an HTLV-I tax sequence in the peripheral blood. Other sets of oligonucleotide primers for HTLV-I gag, pol, env and the long terminal repeat regions and for the HTLV-II pol region were negative in the DNA of the cells. Although patients with T-PLL have been reported to be seronegative for HTLV-I, our findings point to the possibility that HTLV-I infection might be involved in the aetiology of at least some cases of T-PLL and that there may be alternative mechanisms involved in HTLV-associated leukaemogenesis.

The effects of Laserthermia plus chemotherapy on a human gastric cancer transplanted into nude mice was investigated. Three different treatment regimes were tested. The relative tumor growth rate according to the Battele Columbus Laboratories protocol, was as follows. 5-FU chemotherapy, 54%; Laserthermia, 31%; and combined treatment with 5-FU and Laserthermia, 21%. The greatest degree of tumor regression was achieved in the combined therapy group. It is suggested that the combination therapy of Laserthermia plus antitumor agents may be useful in the treatment of gastric cancer in humans.

A cDNA clone of silkworm (Bombyx mori) larval hemolymph antitrypsin (sw-AT) has been isolated from a fat body cDNA library. The cDNA has an open reading frame which codes a 392-amino acid residue polypeptide comprising a 16-residue signal peptide and a 376-residue mature sw-AT of Mr 41,805. The reactive site of sw-AT for inhibition of bovine trypsin [Sasaki, T. et al. (1987) J. Biochem. 102, 433-441] was identified as Lys343-Val344. Alignment of the sw-AT amino acid sequence with those of 11 members of the serpin superfamily of proteins clearly confirmed the homology of sw-AT with serpins. The amino acid sequence of sw-AT is 56% identical with that of the proteinase inhibitor from a lepidopteron, Manduca sexta [Kanost, M.R. et al. (1989) J. Biol. Chem. 264, 965-972], but the sequence around the reactive site shows no homology and the inhibitory specificity for proteinases is very different.

A treatment to evaluate numerically both the direct and exchange amplitudes within the exact framework of the eikonal approximation is considered. The eikonal or unrestricted Glauber-scattering amplitude is decomposed into two partial amplitudes. One of them is reduced to either a closed form for the direct amplitude or to a single-integral expression for the exchange amplitude and is found to approach the Born amplitude at high energy. The other amplitude is expressed in a double-integral form in which the numerical convergence is clearly recognised. By evaluating the direct and exchange amplitudes the results for electron-hydrogen elastic scattering are obtained without the difficulty related to numerical convergence.

Etude sur les alliages Ni x Zr 1−x avec 0,18≤x≤0,90. Il semble exister un ordre chimique a courte distance resultant en une forte correlation de faire Zr-Ni et une faible de plus proches voisins Ni-Ni. Pour les alliages ferromagnetiques amorphes Fe-B, la diffraction de neutrons polarises donne des informations sur les structures partielles

Most cases of infantile anal fistula are treated conservatively, because some of the cases heal spontaneously and frequent reccurrences are observed even after operation.In recent year, the author have witnessed good results of positive operation for this diease. In this paper summerized the results of operated 100 cases in recent 7 years at our clinic.Only few cases showed reccurrence. The period of illness could be shortend by the operation.In conclusion, positive operation for the infantile anal fistula should be tried.

The importance of digital examination for the diagnosis of anal diseases was emphasized. Zeroid, a plastic cylinder 1.2 cm in diameter 10 cm in height with cooled solvent, showed good results for the conservative treatment of anal diseases.As to operative procedures, ligation and excision method was recommended.

It has been considered that incarcerated hemorrhoids should be treated conservatively. But, in our openion, the operation followed by conservative treatments seem to have many weak points, such as local infections and thrombophlibitis. Further, conservative treatment force a patient to bear pain.In this paper, good results of the ligature and excision method for the incarcerated hemorrhoids at their painful stage are presented.No complication after the operation of 239 cases occurred. In conclusion, immediate operation by the ligature and excision method should be tried first.

Isotope shifts in nuclear magnetic shielding have been calculated in the molecular hydrogen system by a semiempirical procedure, which assumes that the high-frequency term varies with internuclear distance R as Rm. It has been found, through a comparison with other investigations, that the choice of m = −1.9 appears to be reasonable. If this choice is taken, then the isotope shift between HD and H2 becomes (0.047 ± 0.02) × 10−6, while the shielding constant for H2 is (26.51 ± 0.3) × 10−6.

Thirty six cases of foreign bodies in the anorectum experienced in recent 6 years were investigated.1) Male and female were in the ratio 25 : 11. Average age of the patients was 51 : 4. These foreign bodies have tendency to be found in older patients.2) The foreign bodies by oral route and by anal route were analysed. Thirty three cases were by oral route and 3 cases were by anal route. Among the foreign bodies by oral route, 19 cases were bone from fish. An whisky glass, chopsticks and a safety pin were found in the rectum inserted by anal route.3) Complications caused by the foreign bodies were abscess formation in the anal region, passage disturbances of feces and urinary retention.4) All of these foregn bodies could be removed per anum. Twenty seven cases needed anesthesia. Operative removing were performed among 23 cases.

Recently, the muonic three-body systemdtμ, has aroused considerable attention in relation to the realization of a useful muon-catalyzed fusion [1, 2]. This bound system must be solved up to 6 digits in energy to establish the muon catalyzed fusion process. We applied the ATMS method [3] and the coupled-channel method [4] to the Coulomb three-body system and obtained the detailed information on the energy levels and the wave functions of the bound states. Further we investigated the effect of the nuclear interaction on the fusion rate in the bound states [5] and developed a formalism to calculate the muon sticking to4He [6]. In this paper we report the results obtained in our collaborations: Figure 2 shows up our main results.