bluuebunny/crossref_metadata_2025_split huggingface dataset:
Anti-Drug Antibody Responses to Lumiliximab Are Not Detected in Relapsed Refractory CLL Patients Treated with Lumiliximab in Combination with FCR in a Phase 1/II Study
Authors: Ranuio Julie , Estrellado Annalee , Harris Sarah , Shannon Megan , Subramanyam Meena , Hughes Steve , Woodworth James , Tangri Shabnam 
Published: April 2020

Abstract
Lumiliximab, an anti-CD23 monoclonal antibody that binds specifically to human CD23, is under investigation for the treatment of patients with relapsed refractory chronic lymphocytic leukemia (CLL). The clinical safety monitoring for Lumiliximab includes testing for the presence of anti-drug antibodies (ADAs) in patient serum samples. In order to determine if patients develop an ADA response to Lumiliximab, serum samples were collected at specific time points throughout a Phase I/II study (152–30) of Lumiliximab in combination with FCR in relapsed CLL. Patient sera from this Phase I/II study were tested in a standard bridging ELISA in which samples were added to a drug coated plate and ADAs were detected using conjugated drug and colorimetric detection. Validation results showed the sensitivity of the assay was 1000ng/ml using a purified polyclonal rhesus anti-Lumiliximab antibody in undiluted human sera. ADAs were not detected in any patients that had evaluable post treatment (Week 13, Week 25, Week 41, and Month 12 post first dose) serum samples in the 152–30 study. Over the last few years, there has been an effort in the immunogenicity field to develop ADA assays with increased sensitivity and to adapt formats that, in general, have increased drug tolerance, i.e. reduction of drug interference in the assay. Therefore, a new ADA assay was developed using Meso Scale Discovery (MSD) technology that has become the reliable and current method for detection of ADA responses and incorporates an acid dissociation step that has been shown in literature to improve drug tolerance. Using the new method the same evaluable samples from the phase I/II study were tested to determine if any previously unseen ADA responses could be detected. The samples were incubated with biotinylated and ruthenylated drug, added to a streptavidin coated plate and ADAs were detected using electrochemiluminescence. Validation results show that the newly developed MSD assay has sensitivity of 250ng/ml using a purified polyclonal chicken anti-Lumiliximab antibody in undiluted human sera. All the samples tested in the MSD assay (n=32) were negative despite greater sensitivity than the ELISA method and an acceptable drug tolerance at the time points samples were drawn. PK concentrations at the specified ADA collection times ranged between 0μg/ml and 114μg/ml whereas drug tolerance ranges from 1μg/ml to 200μg/ml depending on ADA concentrations. Drug interference is a major factor in not being able to detect antidrug responses, yet several of the corresponding post dose PK time points showed low to no drug levels and yet no anti-drug antibodies were detected. These data imply that Lumiliximab when administered in combination with FCR has a low immunogenicity rate.
Do Microenvironmental Factors Play Important Role in Supporting the Maintenance of Leukemic Stem Cells of Chronic Myelogenous Leukemia?
Authors: Cho Yeon Hee , Kim Dong Hwan , Jang Jun Ho , Kim Seok Jin , Kim Kihyun , Kim Won Seog , Jung Chul Won 
Published: April 2020

Abstract
Background : In the era of tyrosine kinase inhibitor(TKI) for chronic myelogenou leukemia(CML), it is becoming more important to develop the way to get rid of the leukemic stem cells(LSC), as there is frequent relapse when TKI therapy is stopped even when molecular remission is achieved. Some researchers reported macrophages and Wnt signaling in bone marrow microenvironment might play significant role in the development and the progression of CML.
Methods : We isolated CD34+CD38− LSC from bone marrow cells from 8 patients with CML chronic phase and also CD34+CD38− normal hematopoietic stem cell from 7 normal controls using magnetic cell separation system. CD34+CD38− cells were cultured on stromal cells for 5 weeks and transfered to methylcellulose media for long-term culture initiating cell(LTC-IC) assay. We checked whether there is a difference in LTC-IC capacity of LSC when cultured on leukemic or normal stromal feeder cells. CD45− stromal supporting cells were established by 3-week culture after 12Gy irradiation of CD34− bone marrow cells. We studied the expression of VCAM, ICAM, E-selectin in stromal cells and the presence of BCR-ABL transcript. Wnt and β-catenin, frizzled, Notch1 were analyzed in the LSCs. CD14+ cells were isolated and added in LTC-IC assay whether the presence of leukemic or normal CD14+ cells would make a change in the maintenance of LSCs.
Results : The number of CD34+CD38− cells in CML was in the range of 1.3 × 103 to 9.2 × 104, that we could do only two sets of 2 × 2 LTC-IC assay that statistical analysis was impossible. LTC-IC frequency of LSCs was about 5-times higher compared to that of normal stem cells irrespective of the kind of stromal cells used. CD14+ cell co-culture did not changed significantly the LTC-IC frequency both in normal and LSCs. Wnt 2B, 5A and 10B were highly expressed in LSCs irrespective of the stromal cells. β-catenin was also strongly expressed in LSCs compared to normal stem cells. Notch1 was expressed only in CD34+CD38− LSCs but not in normal stem cells. The kind of stromal cells or the addition of CD14+ cells in LTC-IC assay did not affected the Notch1 expression in LSCs.
Conclusion : CML CD34+CD38− LSCs seem to have more LTC-ICs and wnt and notch pathway may play important role in the maintenance of LSCs that could be therapeutic targets.
Heme Oxygenase-1(HO-1): A Promising Novel Target Molecule for Overcoming Imatinib Resistance in Chronic Myeloid Leukemia
Authors: Wang Jishi 
Published: April 2020

Abstract
Objective : HO-1 is a microsomal enzyme catalyzing the first, rate-limiting step in degradation of heme, HO-1 is a inducible isoform of HO, it can be strongly induced in response to cellular stress and diverse oxidative stimuli, including its substrate heme, Many studies have convincingly shown that HO-1 is a cytoprotective and antiapoptotic enzyme. the objective of this study was to investigate the influence on the K562 cell growth and apoptosis after hemin-induced HO-1 expression, and to investigate the influence on K562 cells and imatinib-resistant CML cells after ZnPPIX-induced HO-1 inhibition.
Methods: different concentrations hemin (0umol/l A20umol/l 30umol/l)was used respectively to induce HO-1 expression of cultured chronic myeloid leukemia cell line K562, then detected HO-1 mRNA expression under different time by RT-PCR, and MTT was used to detected the viability of K562 cells. In addition, we used STI571(2 μmol/L) deal with the hemin-induced cells, then confirm HO-1 protective effect against STI571 use MTT. Then ZnPPIX was used to inhibition HO-1 expression of K562 and imatinib-resistant cells, similarly, RT-PCR and MTT was used for analyzed.
Results:
The HO-1 mRNA was not tested when absence of hemin, 8h after treated with hemin of 20 μmol/L, we can test the HO-1 mRNA expression, and at 16h the expression is reach to the peak, 16h after treated hemin under different concentrations (10umol/l, 20umol/l, 30umol/l), we found the expression is in a dose-dependent manner. In the group of 10 umol/l and 20 umol/l, the survival of cells is significantly increased in comparison to the control and also have significantly difference in the two groups(p&lt;0.05), in the group of 20 μmol/L, 16h to 48h after hemin-induced, the survival of cells presents a time-dependent manner. In the group of 10μmol/L and 20 μmol/L, exposure of K562 cells to STI571 resulted in a substantial decrease of cell viability in comparison to the STI571 single treatment group(p&lt;0.05). ZnPPIX-induced HO-1 inhibition leads to induction of apoptosis in K562 cells, having significant difference with the control group(p&lt;0.05). ZnPPIX-induced HO-1 inhibition can suppress the survival of imatinib-resistant cells(p&lt;0.05).
Conclusion: our studies have shown that hemin-induced HO-1 gene expression may promote the proliferation of K562 cells, and can against the cell apoptosis. And we found hemin-induced HO-1 gene expression can protect K562 cells against STI571-induced apoptosis, ZnPPIX-induced HO-1 inhibition leads to decreased viability of imatinb-resistant CML cells. these all indicates HO-1 may represent a novel targeting in CML.