bluuebunny/crossref_metadata_2025 huggingface dataset:
Good Prognosis of Adult Immune Thrombocytopenia in a Provincial Hospital
Authors: Jutiamornlerd Siriporn , Uaprasert Noppacharn , Rojnuckarin Ponlapat
Published: December 2023

Background: Immune thrombocytopenia (ITP) is an autoimmune disease resulting from platelet destruction and impaired platelet production. Some patients experience relapses or refractoriness after corticosteroid treatment, necessitating second-line treatments. The majority of adult ITP cases become chronic or relapsing. The aim of this study was to assess the clinical course and identify factors related to sustained responses to treatment in patients with ITP.

Methods: This observational retrospective cohort study was conducted from January 2013 to December 2022 at Trang Hospital, Trang Province, Thailand.

Results: A total of 125 newly diagnosed ITP patients were enrolled, with an average follow-up duration of 48.49 ± 41.49 (mean ± SD) months. The average age was 42.34 ± 18.44 years, the average platelet count was 11.66 ± 14.39 x 10 9 /L, and the average platelet recovery time after treatment was 43.14 ± 101.32 days. The mean response duration after the first-line treatment was 25.99 ± 29.39 months. The characteristics of the patients were as follows: age below 60 years (78.4%), female sex (75.2%), pregnancy (1.6%), primary ITP (67.2%), secondary ITP (32.8%), bleeding symptoms (54.4%), platelet count <10 x 10 9 /L (64%), platelet count 10-30 x 10 9 /L (28.8%), platelet count 30-50 x 10 9 /L (5.6%), platelet count >50 x 10 9 /L (1.6%), splenectomy (4%), observation only (2.4%), loss to follow-up (10.4%), and death (4%). The most common cause of secondary ITP was systemic lupus erythematosus (63%). The first-line treatment consisted of dexamethasone (70.4%), prednisolone (23.2%), and/or intravenous immunoglobulin (4%).

The clinical courses were as follows: recovery within 3 months in 68.8%, persistence in 13.6%, and chronic ITP in 17.6%. The sustained responses to corticosteroid treatment at 6 months included complete response (CR) (58.4%) and response (R) (9.6%), as shown in Table 1. Second-line treatments were required in 22.4% of cases, including colchicine (60.7%), azathioprine (35.7%), cyclophosphamide (25%), dapsone (17.8%), vincristine (10.7%), mycophenolate mofetil (7.1%), cyclosporine (3.5%), and/or eltrombopag (3.5%). The sustained responses to second-line treatments at 6 months were CR in 12% and R in 8.8%. Female sex ( P-value 0.007) and primary ITP ( P-value 0.046) were significantly associated with the sustained response (CR+R) to corticosteroids at 6 months. Secondary ITP ( P-value 0.036) was also associated with the sustained response (CR+R) to second-line treatments at 6 months (Table 2).

Conclusions: This study reveals a favorable prognosis for ITP, with a majority of patients showing sustained response (CR+R), which differs from findings in studies conducted at referral centers. Female patients and those with primary ITP are more likely to respond. Further confirmation is needed through a large prospective cohort study in community hospitals.
Gpibα Caar-T Cells Induce Autoreactive B Cells Elimination to Treat Immune Thrombocytopenia
Authors: Zhou Jie , Mei Heng , Xu Yanyan , Shu Jinhui , Huang Linlin , Xu Min , Jiang Haojie , Liu Junling , Hu Yu
Published: December 2023

Background: Immune thrombocytopenia (ITP) is a bleeding disorder caused by autoantibodies against platelet glycoproteins (GPs). A key attempt to treat ITP is to eliminate the autoreactive B cells. Here, we proposed a new strategy for B cell-depletion therapy of ITP, chimeric autoantibody receptors T cells (CAAR-T), which could trap autoreactive B cells and perform specific killing. GPIbα is a significant autoantigen found in ITP patients and indicates a poor response to standard immunosuppressive treatments. We developed GPIbα-based CAARs in this study.

Aim: To precisely eliminate the autoreactive B cells to treat ITP.

Methods: We constructed various lengths of the native GPIbα ectodomain into the ligand-binding domain of a second-generation CAR structure and expressed the CAAR structure on the HEK293 cell surface to verify its conformational epitope. We immunized BALB/C mice with human platelet lysates to obtain anti-GPIbα hybridomas and performed a binding assay to confirm selected hybridoma antibodies' binding ability to GPIbα CAAR. We performed cytotoxicity assays to assess GPIbα CAAR-T-cell selective cytolysis against anti-GPIbα B cells in vitro and developed a xenograft model to assess the cytolytic efficiency and security of GPIbα CAAR-T cells in vivo. A plasma antibody binding assay was used to test GPIbα CAAR-T cells' ability in reaction with native GPIbα autoantibodies from ITP patients. We applied an enzyme-linked immunospot (ELISpot) assay to verify GPIbα CAAR-T cells' potential in eradicating autoreactive B cells of ITP patients.

Results: We successfully expressed GPIbα CAAR structure on the cell surface and proved that the conformational epitope of GPIbα CAAR is consistent with that of the native human platelet GPIbα. The Schematic of GPIbα CAAR structure is presented in Figure 1A. By immunizing BALB/C mice with human platelet lysates, we obtained four anti-GPIbα hybridomas with different CAAR binding sites. Vitro cytotoxicity assays confirmed GPIbα CAAR-T-cell selective cytolysis of anti-GPIbα hybridoma cells. In a hybridoma xenograft model, we verified that the CAAR-T cells could persist and selectively lyse autoreactive B cells and subsequently reduce autoantibody titers without apparent off-target organ toxicity. Through the plasma antibody binding assay, we demonstrated that GPIbα CAAR could react with sera from ITP patients with anti-GPIbα antibodies (Figure 1B). ELISpot analysis showed that GPIbα CAAR-T cells successfully eliminated anti-GPIbα B cells from a patient with refractory ITP, and normal B cells were not destroyed compared to anti-CD19 CAR-T cells.

Conclusion: Our study provides a proof of concept for CAAR-T cell therapy to eradicate autoimmune B cells while sparing healthy B cells, which would compensate for the defects of traditional B cell depletion therapy in ITP. GPIbα CAAR-T therapy would be a promising treatment for refractory and relapsed ITP patients.
Relationship between Sarcopenia and the Heterogeneity and Microenvironment of Acute Myeloid Leukemia Based on Single-Cell Transcriptome Sequencing
Authors: Sun Qian , Liu Wenjie , Hong Ming , Zhu Yu , Li Jianyong , Qian Sixuan
Published: December 2023

Background: High intratumoral heterogeneity heterogeneity (ITH) and changes in bone marrow microenvironment (BMM) components are important for drug resistance and recurrence in patients with acute myeloid leukemia (AML), and may also influence the phenotype of sarcopenia. This study aims to determine the relationship between sarcopenia and the heterogeneity and microenvironment of AML.

Method: Body composition was assessed by bioelectrical impedance analysis before treatment. Sarcopenia was diagnosed by low muscle quantity according to European Working Group on Sarcopenia in Older People 2 (EWGSOP2). Bone marrow samples from 9 patients with relapsed or refractory AML were used to perform single-cell transcriptome sequencing (scRNA-seq) to analyze the differences in tumor cell subsets and microenvironmental components between AML patients with or without sarcopenia. The samples of 19 newly diagnosed AML patients were further collected for bulk RNA-seq, and deconvolution was done. Bioinformatics methods were used to verify the differences in cell subsets of microenvironmental components in patients with and without sarcopenia, as well as the genes and signaling pathways of tumor cells with differential expression.

Results: Nine leukemia-like cell subsets were identified in AML patients by scRNA-seq. In AML patients with sarcopenia, hematopoietic stem cell (HSC) subsets accounted for a significantly higher proportion of malignant cells, while progenitor cells and monocytes showed a higher proportion in patients without sarcopenia. In sarcopenic group, there were significantly fewer neutrophils, megakaryocytes, and CD8 + Tcm cells in the microenvironment component, while significantly more immunosuppressed Treg cells. After deconvolution of bulk RNA-seq data from 19 newly diagnosed AML patients, it was confirmed that Treg cells were more distributed in sarcopenic patients. Among them, 167 genes were significantly up-regulated, and the most significantly up-regulated 5 genes were ME3, UBE2E3, PCSK5, LAMC1 and PTPRF. A total of 224 significantly down-regulated genes were identified, including RNF18B, TNFAIP3, GALNT2, ORM2 and IL1R2. Pathway enrichment of differentially expressed genes was found, including signaling pathways regulating pluripotency of stem cells, Hippo signaling pathway, protein digestion and absorption pathway, etc.

Conclusion: Sarcopenia may be related to immunosuppressive microenvironment and affect the prognosis of AML patients.
First Report of Asymptomatic Solitary High-Grade B-Cell Lymphoma NOS: An Enigmatic Entity
Authors: Bugazia Seif , Gandhi Nakul , Weerakoon Niluka , Kukreja Geetika
Published: December 2023

Background

High-grade B cell lymphoma (HGBL) is a new disease entity introduced by WHO in 2016, including two types: HGBL with a double hit (DH) or triple hit (TH) and HGBL, not otherwise specified (NOS). HGBL DH/TH are defined by genetic rearrangements of specific oncogenes MYC, BCL2 and/or BCL6, while HGBL NOS is defined purely base on morphology in the absence of these genes. HGBL DH/TH is known for its poor prognostication given its propensity to rapidly progress and disseminate, however HGBL NOS remains ill-defined and poorly understood with no tangible literature describing its clinical nature and therapeutic susceptibility.

Case presentation

A 76-year-old post-menopausal woman with no significant PMH presented for evaluation of painless hematuria during a routine office visit. Evaluation with imaging showed a suspicious incidental liver mass concerning for possible metastatic or primary liver cancer. Obtained labs were unremarkable, including serum LDH, serological testing for hepatitis B/C and HIV1/2, and typical tumor markers. PET scan showed focal uptake of tracer in liver with no nodal involvement. Liver biopsy revealed diffuse infiltration by atypical lymphocytes. Immunohistochemical analysis showed CD20 and CD10 positivity, negative CD30, and high Ki-67 proliferation rate of 100%, however the absence of MYC, BCL6, BCL2, and t(8;14) rearrangements on FISH ruled out Burkitt and DH/TH lymphoma, consistent with HGBL-NOS. Following discussions with Henry Ford Lymphoma Tumor Board, patient was treated with six cycles of R-CHOP, where follow-up imaging indicated remission of her lymphoma.

Conclusion

This represents an extraordinary first report of HGBL NOS manifesting as an isolated primary hepatic lymphoma in the complete absence of clinical symptoms. The significance of this case lies in its atypical extra-nodal site of involvement and total absence of clinical symptoms and serological markers, underscoring the unfulfilled need to further describe this enigmatic entity.
Complete Blood Count Reporting Among Hospitals in the United States
Authors: Go Lewis T , Go Lucas T , Gunaratne Madugodaralalaage DSK , Abeykoon Jithma Prasad
Published: December 2023

Background

Complete blood count (CBC) is the most common test performed in hematology-oncology. A typical CBC report contains over 10 elements. Simplifying the report by removing nonessential elements may benefit clinicians and patients. Little is known about what elements are reported by various hospitals in the US. The objective of our study was to describe how hospitals in the US report CBC.

Methods

After approval from the Mayo Clinic institutional review board, we obtained hospital CBC reported elements from the Epic electronic medical records Care Everywhere health information exchange. CBCs from 2020-2023 were included. For each hospital, we determined the city and state where it's located. We obtained the number and type of elements reported in the CBC and compared the findings between academic and community hospitals.

Results

We collected CBC data from 139 hospitals across 43 states and 102 cities (Figure 1). Fifty-four percent were community hospitals while 46% were academic hospitals. The median number of elements reported by all hospitals was 21 (range, 12-24). This was not different (P=0.98) between academic (median 22 [range, 12-24]) and community hospitals (median 21 [range, 14-24]; Figure 2). The distribution of hospitals according to the number of CBC elements reported were as follows: 12-15 (6%), 16-19 (13%), 20 (25%), 21(6%), 22 (18%), 23 (19%), and 24 (13%). A minority of the hospitals did not report % white blood cell differential count (9.4%) or mean platelet volume (20.9%). Nucleated red blood cell and immature granulocyte count were reported frequently, 46.0% and 56.8%, respectively. However, the frequency of reporting these four elements were similar between academic and community hospitals.

Conclusions

We found substantial variations in CBC reporting among US hospitals even within academic and community hospitals. Most hospitals reported 20 or more elements, twice as many as the hospitals reporting the least number of elements. There is an opportunity to simplify and standardize CBC reporting, which will minimize physician burnout and distraction as well as decrease patient concern and query.
Interactive Pathology Tutorial in Neoplastic Hematology Disorders for Medical Hematology-Oncology Fellows
Authors: Alkhasawneh Ahmad N , Baskovich Brett , Gopinath Arun , Allan Robert W. , Mohamed Abdullah , Quan Walter JR
Published: December 2023

Context: Hematology-Oncology (HO) fellows receive limited instruction in the process of establishing a diagnosis for hematologic neoplasms. Learning neoplastic hematology often occurs in limited encounters such as reviewing slides with pathologists or during a tumor board where the diagnosis has already been established. In the current study, we developed a web-based interactive pathology tutorial in neoplastic hematology disorders for HO fellows to work up simulated cases and establish the diagnosis.

Design: An online system (“Pathology Playground”) was utilized to load case materials including microscopic images and ancillary studies. Twelve high-yield simulated cases of common leukemias and lymphomas likely to be encountered in the daily practice, such as acute promyelocytic leukemia, were included (www.drdoubleb.com/crowdunknowns/study-set/30). At the beginning of each case, trainees review the clinical history and slide images (image 1), and then they are given the option to request additional work up, such as flow cytometry, immunohistochemistry, karyotype and/or molecular studies. Based on the results, they can enter their diagnostic impression. If the diagnosis is correct, the user is shown a short educational presentation about the entity with the key pathology findings.

Results: The tutorial was integrated in the educational curriculum of our HO fellowship program, and bimonthly teaching sessions were held to review two cases each time. During the sessions, trainees request ancillary studies to complete the diagnostic work-up using the software, and interpret the findings. As the case is being worked up by the trainee, the hematopathologists and HO fellowship program director discuss the requested work up findings, the appropriate work-up tools of the case as well as the clinical picture and the implications on the patient management. All of our six HO fellows attended the sessions, and a survey from the trainees showed high ease of use of the system and they viewed it as a very useful educational tool.

Conclusions: The interactive tutorial in hematopathology was highly received by our HO fellows, and they found it beneficial for their education. Expanding the use of this online interactive tutorial and incorporating additional cases would enhance its value as a learning resource.
Leukostasis Twist in Mantle Cell Lymphoma: Systematic Review and Case Report
Authors: Chiu Jodi , Crowther Mark A.
Published: December 2023

Introduction:

Mantle cell lymphoma (MCL) is an aggressive B-cell non-Hodgkin lymphoma. Patients often present with lymphadenopathy, early satiety, and B-symptoms. Despite up to 92% of MCL patients having peripheral blood involvement by flow cytometry, presentation with leukemic involvement and hyperleukocytosis are uncommon.

Hyperleukocytosis, marked by a white blood count (WBC) >100 x 10 9/L, often leads to symptomatic leukostasis due to tissue hypoxia, generally found in acute leukemias. We conducted a systematic review of leukostasis in MCL, and present a case we encountered in our clinical practice of a non-nodal MCL variant showing leukostasis, spontaneous tumour lysis syndrome (TLS), and severe anemia to identify treatment strategies and outcomes.

Methods:

We searched PubMed for peer-reviewed articles and abstracts on “mantle cell lymphoma,” “hyperleukocytosis,” and “leukostasis.” All articles and abstracts that discuss hyperleukocytosis leading to leukostasis in MCL were included.

Results:

The search yielded eight results. After exclusions, five case reports and abstracts were included. These studies featured different MCL variants, presenting WBCs ranging from 121 to 1227 x 10 9/L, and various leukostasis symptoms. Leukapheresis was used in all five published cases, leading to lower WBCs and symptom improvement. Results are summarized in Table 1.

Our patient was found to have the small lymphocyte variant of MCL, with WBC of 543.3 x 10 9/L on presentation. Our facility does not have leukapheresis capability. As a result, our initial treatment consisted of only supportive care and escalating doses of corticosteroids, with close monitoring for TLS. To our knowledge, this report is the first demonstration that IV methylprednisolone, without leukapheresis, is effective for acute management of leukostasis. Our patient's WBC reduced from 543.3 x 10 9/L to 230 x 10 9/L, leading to improved neurological function and oxygenation. Steroids were particularly useful in our patient with secondary, MCL-related warm autoimmune hemolytic anemia.

Conclusion:

This review highlights the importance of recognizing non-specific leukostasis symptoms, even in patients with mature B-cell lymphoproliferative disorders. Leukapheresis is effective in treating leukostasis in MCL patients. In hospitals without leukapheresis, escalating doses of IV methylprednisolone can be considered as a safe and effective strategy.
Opoid Use Disorder and Vaso-Occlusive Crisis Outcomes: A Retrospective Assesement
Authors: Sanni Abdulsabur , Goble Spencer , Linzer Mark
Published: December 2023

Introduction

Concerns by providers for addiction can lead to under-treatment of pain in patients with sickle cell disease presenting with vaso-occlusive crisis. In patients who experience frequent vaso-occlusive crises, opioid dependence may be due to the need for pain control as opposed to addiction and the clinical meaning of a diagnosis of opioid use disorder and its impact on outcomes in this population is not known.

Methods

We retrospectively assessed hospitalizations in the United States for vaso-occlusive crisis in adult patients afflicted with Sickle cell disease from 2016-2019 using the National Inpatient Sample database. Outcomes were compared between those with and without a diagnosis of opioid use disorder using linear and logistical regression.

Results

In total, there were 273,460 hospitalizations for vaso-occlusive crisis, 23,120 (8.5%) of which involved a secondary diagnosis of opioid use disorder. Black Race was associated with opioid use disorder, but age, sex and income were not. After adjusting for confounders, hospital length of stay was increased in patients with opioid use disorder by 1.23 days (95% CI: 1.02 to 1.45, p<0.001) and total hospital charges were increased by $5,566 (95% CI: 3,863 to 7,269, p<0.001) (Table 1). Mortality was decreased in vaso-occulsive crisis hospitalizations in those with opioid use disorder, but the difference was not significant (OR=0.58, 95% CI: 0.026-1.33, p=0.199).

Conclusion

Opioid use disorder is associated with an increased length of stay and cost in patients with vaso-occlusive crisis. Providers should consider under-treatment of pain as a potential diagnosis when addiction concerns are present and individualized pain plans should be explored.
The Concordance between Laboratory and POCT INR and the Value of Bedside Measurement of INR in Patients on Warfarin Undergoing Dental Procedures: A Prospective Cohort Study
Authors: Albarkheel Abdullah , Owaidah Tarek M , Tamim Hani , Alahmadi Abdullah , Sheikh Aamir , Mushtaq Ali
Published: December 2023

Introduction: Patients on anticoagulation particularly warfarin suffers when they have a dental problem that requires intervention. Most of dentists worry and refuse to perform procedures on patients on warfarin because of the risk of bleeding and request the availability of international normalized ratio (INR) results before procedure which can delay the procedure and add another burden on the patient particularly who live away from the treating center.

Objective: To evaluate the efficacy and the value of using point of care testing (POCT) (INR) in patients on warfarin undergoing dental procedures. We also had analyzed the frequency and severity of dental bleeding in patients on warfarin therapy and the concordance between Laboratory based INR and POCT INR. This is the first interim analysis on ongoing prospective study.

Methods: A cohort study of 30 patients on chronic use of anticoagulation therapy with warfarin for different clinical indications who planned to undergo dental procedures at a teaching hospital in Saudi Arabia. The patients' demographic and clinical characteristics, type and severity of bleeding, and INR measured by both HemoChrone (POCT) and prothrombin time (PT) were performed at same time. Descriptive statistics, paired samples statistics, correlations, and analysis of variance (ANOVA) were performed.

Results: The sample consisted of 60% females, with a mean age of 49.9 (SD 13.3) years. The most common indications for warfarin therapy were mechanical aortic valve replacement (56.7%), atrial fibrillation (13.3%), and atrial flutter (16.7%). Before the procedure, the mean Laboratory INR was 2.6 (SD 0.6) and the mean HemoChrone POCT INR was 2.8 (SD 0.8). There was a significant positive correlation between the two methods (r = 0.81, p < 0.001). There was no significant difference in the INR and type of bleeding, dentist performed the procedure, or dose of warfarin. There was no correlation between the Bleeding severity or type and the INR level in patients undergoing different dental procedures. Even among those with HemoChrone POCT INR > 3, which is considered high and increases the risk of bleeding, there was no significant correlation with bleeding.

Conclusion: These results imply that warfarin therapy can be safely continued for most dental procedures without increasing the risk of bleeding regardless of the INR results. The HemoChrone POCT INR is a dependable and practical method to monitor the effect of warfarin in dental settings which can reassure the dentists before procedure.

Figure 1: The concordance between Laboratory and POCT INR
BOD1L Facilitates Chromatin Binding of SETD1A and Promotes Leukemia Cell Growth
Authors: Hoshii Takayuki , Kikuchi Sota , Shibata Seito , Matsumoto Makoto , Rahmutulla Bahityar , Fukuyo Masaki , Armstrong Scott A , Kaneda Atsushi
Published: December 2023

The H3K4 methyltransferase SETD1A plays an essential role in leukemia through a non-catalytic activity which regulates transcriptional pause release of RNA polymerase II. SETD1A localizes to the transcriptional start site of numerous actively transcribed genes, but primarily regulates the expression of a restricted set of genes within the DNA damage response as well as heme biosynthesis pathway. However, the roles of upstream regulators or essential components of SETD1A chromatin binding remain unclear.

Using the DepMap database and a CRISPR-tiling screen method, we found that BOD1L is the most correlated co-dependency with SETD1A in human cancer cell lines and that the Shg1 domain of BOD1L plays an indispensable role in human MOLM-13 leukemia cell line. BOD1L knockout increases apoptosis and reduces cell cycle. The strong BOD1L dependency in AML cells was also confirmed using a mouse MLL-r leukemia model in vitro and in vivo. SETD1A catalytic domain mutant AML cells still responded to the BOD1L sgRNAs that suggests the role of BOD1L is independent from the SETD1A catalytic function. Importantly, BOD1L knockout mimics the transcriptional profiles of SETD1A knockout cells and reduces the expression of genes in DNA damage response and heme biosynthesis. BOD1L loss immediately reduces SETD1A distribution on chromatin and increased the NELFE and RNA polymerase II signals at transcriptional start sites. These results show that BOD1L is an essential component of the non-catalytic SETD1A function in transcriptional pause release.

We also performed an immunoprecipitation assay using tagged constructs to examine the interaction between BOD1L and SETD1A. Our data indicate that SETD1A associates with BOD1L through the FLOS domain, especially in the C-terminal region. In addition, we found that the loop structure in the Shg1 domain of BOD1L is associated with the nonenzymatic domain of SETD1A. A split luciferase complementation assay reveals that the BOD1L Shg1 domain and SETD1A FLOS domain interact. To demonstrate the role of BOD1L chromatin interaction, we enforced localization of the Shg1 domain fragment at a silent gene locus and it successfully induced ectopic SETD1A recruitment to chromatin and formation of the COMPASS complex. However, we observed neither induced H3K4me3 modification nor RNA polymerase II recruitment. Taken together, while the BOD1L-SETD1A complex is not sufficient to act as a pioneer complex for active transcription, our study identified that BOD1L facilitates chromatin binding of SETD1A through the Shg1-FLOS domain interaction and then regulates transcriptional pause release, and that this interaction is a potential therapeutic opportunity for SETD1A-targeting therapy.
Obtaining M Protein Immune Reconstitution As Soon As Possible after Tandem Autologous Hematopoietic Stem Cell Transplantation Improves the Prognosis of Patients with Newly Diagnosis Multiple Myeloma
Authors: Zhu Huihui , Li Juan
Published: December 2023

Objective: In the era of new drug therapy, autologous hematopoietic stem cell transplantation after induction therapy is the standard of care for patients with multiple myeloma, and Tandem autologous hematopoietic stem cell transplantation is a treatment option to improve the prognosis of patients with high-risk cytogenetics, and obtaining M protein immune reconstitution as soon as possible after double transplantation can further improve the survival of such patients.

METHODS: A total of 12 patients with newly diagnosed multiple myeloma diagnosed from June 1, 2019 to April 30, 2021 at the First Affiliated Hospital of Sun Yat-sen University were included, and all patients underwent tandem autologous hematopoietic stem cell transplantation after induction therapy containing bortezomib, and the prognostic survival of patients who obtained M-protein reconstitution after transplantation was analyzed

Results: 58.3% (7/12) of patients obtained M protein immune reconstitution after Tandem transplantation, the most common type of M protein was IgG-type M protein, the median time to obtain M protein immune reconstitution was 3 months (2-9 months) after Tandem transplantation, the median duration of reconstitution was 3 months (3-6 months), and all patients were R-ISS stage II-III, 83.3% (10/12) of patients had high-risk cytogenetics, all patients achieved at least VGPR efficacy after transplantation, with a higher CR rate in the M protein immune reconstitution group (71.4% vs 60.0%, P=0.679), although not statistically significant, and a median survival time of 2 years in the immune reconstitution group

CONCLUSION: Tandem transplantation is a treatment option to improve prognosis in patients with high-risk cytogenetic multiple myeloma, and patients who received M protein immune reconstitution after Tandem transplantation significantly improved survival time.
Impact of Psychiatric Comorbidity on Readmissions of Sickle Cell Disease -a Nationwide Readmissions Database Study
Authors: Kamal Syeda Ashna Fatima , Jhu Joshua , Maiocco Giuseppe , Prakash Vivek , Hussain Mohammad Junaid , Azhar Waqas , Agarwal Ishan , Bloch Evan , Tobian Aaron , Goel Ruchika
Published: December 2023

Introduction:

While advances in the management of sickle cell disease (SCD) treatment have improved outcomes, readmission rates among patients remain a significant concern. A previous study with the National Readmission Database (NRD) 2016 found that alcohol use, tobacco use, opoid use ,substance use and depression were associated with SCD readmission even though the general rate of readmission has decreased over the years. This study aims to update the literature, and further determine the predictors associated with increased readmission among individuals with SCD.

Methods:

Using the Nationwide Readmissions Database (NRD), we evaluated readmissions for SCD for the year 2016-2020. On the basis of International Classification of Diseases, TenthRevision, Clinical Modification (ICD-10-CM) codes, we identified SCD hospitalizations (D57.0, D57.1, D57.2, D57.4, D57.8), and comorbid conditions. Usingamultivariateregressionanalysis,weestimatedtheindependent predictors of 30-day readmissions of SCD. We used STATA® Version 18.0 Software (STATACORP, TEXAS, USA) for analysis. The p-value was set at p < 0.05 for statistical significance.

Results:

A total of 262,571 index admissions for adult patients with a primary diagnosis of SCD were admitted within the study period; 61,687 patients (23.6%) were readmitted within 30 days. A total of 886 and 105 deaths were reported respectively among index admissions and readmissions. Compared to index admissions, readmissions had decreased THC of 2147 U.S. dollars (95%CI: -2678 - -1615), while we found no significant change in the LOS between index admissions and readmissions. Common causes of increased rate of readmissions were SCD with mood disorder (HR=1.15, 95%CI: 1.11-1.20) , anxiety disorder (HR= 1.16, 95%CI: 1.11-1.20) and opioid use (HR=1.42, 95%CI: 1.35-1.50). Conditions associated with lower rates of readmission are Alcohol use disorder (HR 0.76, 95%CI: 0.62-0.94) and cannabis use ( HR=0.94, 95%CI: 0.88-0.99).

Discussion:

Our findings suggest that among individuals admitted with a primary diagnosis of SCD, having concurrent mood and anxiety disorders, and opioid use disorder increases rates of readmission. Interestingly, we observed that alcohol use disorder and cannabis use was associated with lower rates of readmission in SCD patients. Optimizing the comorbid conditions in patients with SCD may reduce readmission rates. However, further studies are required to establish the relationship between psychiatric comorbidity and healthcare resources utilization among SCD patients.
Integrative Analysis of the Tumor and Microenvironment to Model the Molecular Heterogeneity Underlying the Response to Cevostamab in Relapsed/Refractory Multiple Myeloma
Authors: Hamidi Habib , Nakamura Rin , Roy Sumi , Rajasekaran Kamalakannan , Diaz Giovanni , Dos Santos Cedric , Sumiyoshi Teiko
Published: December 2023

Background: Cevostamab is an FcRH5xCD3 bispecific antibody that facilitates T-cell directed killing of myeloma cells and has shown promising activity and a favorable safety profile as monotherapy in patients with relapsed/refractory multiple myeloma (RRMM) in the ongoing Phase I GO39775 trial (NCT03275103; Trudel et al. ASH 2021). We performed an integrative analysis of peripheral blood (PB) and bone marrow aspirate (BMA) samples collected at baseline from patients in GO39775 to understand the heterogeneity of patients with RRMM and potential correlations with clinical outcome.

Methods: PB and BMA samples were collected at baseline from 100 patients in GO39775. Patients who had less than 25% missingness in any data layer were selected from the single-step (3.6/90-252mg dose levels [n=49] and 3.6/90mg tocilizumab premedication group [n=14; Trudel et al. ASH 2022]) and double-step (0.3-1.2/3.6-7.2/60-160mg dose levels [n=37]) dose-escalation and dose-expansion cohorts. To characterize immune cells, samples were profiled at a central laboratory by 8-color flow cytometry. Cytokine profiling was performed on baseline plasma samples via electrochemiluminescence and digital ELISA. Serum B-cell maturation antigen (sBCMA) levels were quantified using hybrid immunoaffinity capture by liquid chromatography with tandem mass spectrometry. For the unbiased integrative analysis, a Similarity Network Fusion algorithm was applied to preprocessed flow cytometry data from PB and BMA samples, and to baseline characteristics including demographics, prior therapies, and selected clinical laboratory analytes. Data were imputed in samples with less than 25% missingness using a random forest algorithm.

Results: Initial findings from unsupervised machine learning-based integrative clustering analysis of baseline samples from multiple immunological and clinical data types revealed three patient subpopulations with unique biological features, illustrating the molecular heterogeneity of patients with RRMM. Each cluster was characterized by distinct patterns of immune cell proportions and activation states. Interestingly, these immunologically defined clusters also corresponded to clinical outcomes. Cluster 1 had the strongest response to cevostamab, followed by Cluster 2 and Cluster 3.

Conclusions: Our novel, unbiased, machine learning, integrative clustering approach not only revealed unique molecular subtypes that illustrate the heterogeneity of patients with RRMM, but also identified immunological features associated with potentially reduced response to cevostamab therapy, which could provide direction for future research.
TRPM8 Contributes to Early Temperature-Induced Platelet Activation
Authors: Stratiievska Anastasiia , Filippova Olga , Ozpolat Hasan Tahsin , Byrne Daire , Bailey S. Lawrence , Mollica Molly , Harris Jeffrey , Esancy Kali , Daka Ajay , Sniadecki Nathan , Lopez Jose A. , Stolla Moritz
Published: December 2023

Background: Like sensory neurons, platelets are activated by a decrease in temperature. However, the molecular mechanism of this temperature-sensing ability is unknown. Platelet activation by temperature could contribute to numerous clinical observations, most importantly to the well-known reduced lifespan of ex vivo-stored platelets for transfusion. In this study, we sought to dissect the molecular basis of the cold-induced platelet activation response and specifically, evaluate the role of the thermosensitive ion channel transient receptor potential cation channel subfamily member 8 (TRPM8) in platelets and their precursor cells.

Methods: We tested for the presence of TRPM8 on the megakaryocytic cell line MEG-01 and human platelets by immunoblotting, confocal microscopy, and imaging flow cytometry. The effect of TRPM8 on platelets was evaluated using the commercially available TRPM8 inhibitor PF-05105679. In addition, we used TRPM8 agonists such as menthol, WS-12, and icilin. Platelet shape change and αIIbβ3 integrin activation were assessed by imaging flow cytometry and conventional flow cytometry. We tested for platelet aggregation using light transmission aggregometry and for intracellular calcium by reading fluorescence on a plate reader after loading platelets with Fura-2-AM. Calcium release in response to temperature decrease was assessed after loading platelets or TRPM8-transfected HEK293T/17 and reading fluorescence in a Real-Time PCR Detection System.

Results: We detected TRPM8 mRNA and protein in MEG-01 cells and platelets but there was a remarkable variability among donors. Further evidence for TRPM8's presence in platelets came from the fact that inhibitors of TRPM8 prevented early temperature-induced platelet activation events, such as αIIbβ3 integrin activation and shape change. Unlike temperature changes, chemical agonists of TRPM8 did not seem to have an acute effect on platelets. Platelets exposed to below-normal body temperature displayed a cytosolic calcium increase, but this was independent of TRPM8 and entirely dependent on the calcium release from the endoplasmic reticulum.

Conclusion: We found evidence for TRPM8 expression in platelets but the expression varies greatly between individuals and is of unclear physiologic significance. Our study suggests that the cold response of platelets is complex, and TRPM8 appears to play a calcium-independent role in the early temperature-induced activation of platelets, while other mechanisms likely contribute to later stages of temperature-mediated platelet response. Because of the high interindividual variability of TRPM8 expression, a population-based approach should be the focus of future studies.
Integrative Genomic and Transcriptomic Analysis Reveals Genetic Alterations Associated with the Early Progression of Follicular Lymphoma
Authors: Gao Fenghua , Liu Hengqi , Wang Xianhuo , Yao Yixin , Zhang Huilai
Published: December 2023

Background Follicular lymphoma (FL), the most common indolent lymphoma, is a clinically and genetically heterogeneous disease. However, the prognostic value of driver gene mutations and copy number alterations has not been systematically assessed.

Patients and Methods Here, we analyzed clinical-biological features of 415 FL patients to identify variables associated with disease within 24 months of first-line therapy (POD24). In addition, we applied whole exome sequencing (WES) to identify genomic alterations that predict POD24 based on 102 FL patients. Furthermore, we characterized the cellular and molecular heterogeneity within and across patients through bulk RNA sequencing and single-cell RNA sequencing.

ResultsPOD24 occurred in 21% of evaluable FL patients. To further examine the effect of POD24 on OS, we used a 24-month landmark approach for Kaplan-Meier curve analysis. As expected, POD24 was related to poor OS, and the 3- and 5-year survival probabilities of patients in whom the disease progressed within the first 24 months were 89.2% (95% CI, 84.5-93.9) and 78.6% (95% CI, 70.4-86.8), respectively, compared with 98.5% (95% CI, 97.8-99.2) and 96.2% (95% CI, 95.0-97.4) in patients who were progression free at 24 months, respectively. Moreover, patients without progression at 24 months were more likely to exhibit a CR to front-line treatment (54% vs. 35%, P = 0.001) compared to the POD24 cohort. Patients with B symptoms, elevated lactate dehydrogenase and β2-microglobulin levels, unfavorable baseline haemoglobin levels, advanced stage, and high-risk FL International Prognostic Index (FLIPI) scores had an increased risk of POD24, with FLIPI being the most important factor in logistic regression. HIST1H1D, known as a driver mutation, was correlated with POD24. Gains of 6q22.2 ( HIST1H1D) and 18q21.33 ( BCL2) and loss of 1p36.13 ( NBPF1) predicted POD24 independent of FLIPI. Integration of the four variants led to the identification of 76% of POD24 patients. Gene expression profiling of FL samples showed that the POD24 cohort was significantly enriched in the inflammatory response (mediated by interferon and tumor necrosis factor), cell cycle regulation (transcription, replication and proliferation) sets and PI3K-AKT-mTOR signaling. This result was further validated with transcriptome-wide information provided by RNA-seq at single-cell resolution.

ConclusionOur study, performed on a large cohort of FL patients, highlights the importance of distinctive genetic alterations and gene expression relevant to disease diagnosis and early progression.
Epigenetic Modulation Enhances the Therapeutic Potential of All-Trans Retinoic Acid in Acute Myeloid Leukemia
Authors: Szymanski Lukasz , Skopek Rafal , Palusinska Malgorzata , Maslinska-Gromadka Karolina , Kraj Leszek , Schenk Tino , Zelent Arthur
Published: December 2023

Epigenetic dysregulation has been strongly associated with the development and progression of acute myeloid leukemia (AML). While all-trans retinoic acid (ATRA), a derivative of vitamin A, has demonstrated significant anti-cancer effects in acute promyelocytic leukemia (APL-AML), its clinical efficacy in non-APL AMLs has been limited. This limited response can be attributed, in part, to the epigenetic silencing of genes targeted by or involved in the ATRA pathway, rendering them unresponsive to the drug. Consequently, our research aims to overcome the epigenetic barriers in AML by combining ATRA with small molecules that possess epigenetic modulating properties.

To investigate the activation of the retinoic acid pathway in response to ATRA and a library of over 650 small molecules with epigenetic activity, we transduced HEK-293 cells with the p-GreenFire-RARE-Tk-Luc reporter construct. Through the screening process, we have identified 11 compounds that show promise as potential candidates for combination therapy with ATRA in non-APL AML. Notably, when combined with ATRA, two specific compounds, a dual PI3K HDAC inhibitor, and a pan-PKC inhibitor, demonstrated a significant 4-5 fold increase in RARE activity, indicating a synergistic effect.

To determine the impact of selected compounds on the functional level, we evaluated the viability, differentiation, and phosphorylation status of ERK1/2, AKT1, MAPK9, p38, and MAPK9 in various cell lines, including HL-60, KG1a, BMNC, and primary MSC and AML cells. The dual PI3K HDAC inhibitor and a pan-PKC inhibitor were noncytotoxic to HL60, BMNC, and primary MSC. Interestingly, dual PI3K HDAC inhibitor with and without ATRA reduced the viability of KG1α by more than 95%. Moreover, dual PI3K HDAC inhibitor and a pan-PKC inhibitor, when combined with ATRA, lead to a 2-3 fold increase in the differentiation of HL60 cells and a 2-4 fold increase in the differentiation of AML cells ex vivo compared to ATRA in combination with the LSD1 inhibitor TCP (treatment control protocol) and untreated cells from AML subtypes M1, M2, and M4. PI3K HDAC inhibitor combined with ATRA significantly reduced p-MAPK and p-p38 levels while increasing the phosphorylation of p53 in HL-60 cells. Furthermore, RNA sequencing and chromatin profiling by Cleavage Under Targets and Tagmentation (CUT&Tag) were performed to gain additional insights into the molecular mechanisms involved.

Based on obtained results, the combination of ATRA with specific dual PI3K HDAC and pan-PKC inhibitors holds promise as a potential therapeutic approach for AML. Nevertheless, further investigations, including studies utilizing patient-derived xenograft models, are warranted to validate the efficacy and feasibility of this proposed treatment strategy.
Time to Lymphoma Treatment within 24 Months in Watchful Waiting Follicular Lymphoma Defines Patients at High Risk for Progression: A Multicenter Analysis
Authors: Gao Fenghua , Liu Jing , Wang Jiesong , Liu Lihong , Li Zhiming , Song Yuqin , Zhang Xudong , Zhou Hui , Sun Xiuhua , Zhang Wei , Xu Bing , Su Liping , Shujuan Wen , Tao Rong , Bai Ou , Zhang Qingyuan , Zou Liqun , Wang Xianhuo , Zhang Huilai
Published: December 2023

Purpose Follicular lymphoma (FL) is a clinically and molecularly heterogeneous disease, watch and wait (W&W) remains a management therapeutic option in patients with advanced-stage, low-tumor-burden and asymptomatic FL in the rituximab era. We continue to use Groupe d'Etudes des Lymphomes Folliculaires criteria for active disease to initiate therapy. We sought to understand whether time to lymphoma treatment (TLT) after diagnosis in patients who managed by W&W was a factor affecting survival outcomes in FL.

Patients and Methods Between 2008 and 2022, 411 FL patients from 16 institutions in China were managed by W&W strategy, and their TLT was retrospectively evaluated. Patients were further divided into training and validation Cohorts. Logistic regression was used to identify and incorporate independent predictors of early TLT into a model with variable scoring. Model performance was evaluated through the area under the receiver operating characteristic curve (AUC) and goodness-of-fit statistics.

Results After a median follow-up of 46 months, 35 percent of W&W patients experience TLT within 24 months (TLT24) after diagnosis, and the 5-year progression free survival (PFS) rate was significantly lower than that of patients who were treatment-free at 24 months (62.3% vs. 89.5%). In multivariable analysis, five clinical factors were identified as independent predictors of TLT24: stage III-IV, β 2 microglobulin ≥ 3mg/L, lymphocyte-to-monocyte ratio＜3.8, bone marrow involved and spleen enlargement. We calculated risk scores (TLT24PI) for each patient and defined three risk groups: low (0-1 points), intermediate (2 points), or high (3-5 points). Its AUC for TLT24 was 0.761 (95% CI, 0.698 - 0.823) in the development cohort and 0.761 (95% CI, 0.698 - 0.823) in the validation cohort. Risk groups were also associated with PFS ( P<0.001).

Conclusion In patients with FL who initially managed by W&W, TLT within 24 months after diagnosis was associated with poor outcomes. We developed a multivariable model that incorporates clinical and laboratory factors to identify patients at high risk for TLT24, which may be useful to identify candidates for early interventional treatment.
Real-World Study of Adherence to Anticoagulant Treatment Guidelines in Patients with Cancer-Associated Thrombosis (CAT)
Authors: Jain Rahul , Yi Alexander , Li Ang , Vigna Chelsea , Khder Yasser , Bebi Ted , Galaznik Aaron , Rusli Emelly , Bloomfield Daniel
Published: December 2023

Background and Objectives:

Clinical guidelines recommend using anticoagulant therapy for 3-6 months to treat cancer-associated thrombosis (CAT). Limited data are available on real-world compliance with these guidelines in the US. This analysis aimed to describe treatment patterns for CAT in the US.

Methods:

A retrospective cohort study on adult patients with active cancer who developed CAT between January 2018 to December 2019 was conducted using integrated closed medical and pharmacy claims data from HelthVerity PS 20 database. Patients were categorized as treated cohort if they initiated anticoagulant or inferior vena cava filter within 15 days prior or 45 days after the first CAT event. Patients were required to have a continuous enrollment of ≥ 12 months prior and ≥45 days after the first CAT date. Clinical characteristics during the baseline period and treatment patterns during the follow-up period were described.

Results:

A total of 2,119 patients met the inclusion and exclusion criteria, with 1,536 (72.5%) patients assigned to the treated cohort (Quan-Charlson comorbidity index [QCI] 2.11±2.0; HAS-BLED score 1.91±1.1 and VTE-BLEED risk score 3.65±1.3). Among the treated cohort, 43.9% initiated treatment with a DOAC; the median treatment duration was 164 days. Of the treated patients, 33.8% had <91 days of treatment duration. Relative to patients with ≥91 days of treatment, patients with <91 days of treatment had a higher comorbidity burden [QCI 2.43±2.2 vs. 1.88±1.9; p-value <0.001], a higher HAS-BLED score [2.07±1.1 vs. 1.8±1.0; p-value <0.001], and higher average VTE-BLEED risk score [3.82±1.3 vs. 3.54±1.2; p-value <0.001].

Conclusion:

Compliance with clinical guidelines on the use of anticoagulation in CAT is suboptimal. Patients who receive <91 days of treatment are more likely to have higher risk features than those who receive ≥91 days of treatment.