bluuebunny/crossref_metadata_2025_split huggingface dataset:
Efficacy and Safety of the Second CAR-T Therapy in Patients with Refractory/Relapsed Acute B-Cell Lymphoblastic Leukemia
Authors: Liu Sining , Cui Qingya , Li Zheng , Cui Wei , Li Mengyun , Qiu Huiying , Xue Shengli , Chen Suning , Jin Zhengming , Miao Miao , Han Yue , Wang Ying , Zhu Xiaming , Yu Lei , Wu Depei , Tang Xiaowen 
Published: December 2023

Background: Chimeric antigen receptor T (CAR-T) cells therapy has shown significant efficacy in relapsed or refractory B-cell acute lymphoblastic leukemia (R/R B-ALL), but a substantial proportion of patients still relapse after a period of remission. The efficacy of a second CAR-T cell infusion is uncertain. The aim of this study was to explore the efficacy and safety of the second CAR-T therapy in R/R B-ALL patients.
Methods: Between August 2018 and October 2022, 35 patients with R/R B-ALL successfully received the second CAR-T treatment. Among them, 18 patients received single CD19 or CD22 CAR-T therapy, 14 patients received tandem CD19/CD22 CAR-T therapy, and 3 patients received sequential CD19 and CD22 CAR-T therapy.
Results: The complete remission (CR) rates were 55.6% (10/18) in patients who received CD19 or CD22 CAR-T therapy, 85.7% (12/14) in patients who received tandem CD19/CD22 CAR-T therapy, and 33.3% (1/3) in patients who received sequential CD19 and CD22 CAR-T therapy (tandem CD19/CD22 vs. others, P=0.045). Tandem CD19/CD22 CAR-T therapy remained one of the significant favorable factors in multivariate logistic regression analysis of CR rate in all patients (hazard ratio, 0.128; 95% CI, 0.017-0.974). A total of 5 patients with severe cytokine release syndrome (Grade ≥ 3) were observed, including 2 patients in the single CD19 CAR-T group, 2 patients in the tandem CD19/CD22 CAR-T group, and 1 patient in the sequential CD19 and CD22 CAR-T group. Of the 23 patients who achieved CR, 4 patients bridged to allogeneic hematopoietic stem cell transplantation (allo-HSCT) and 3 patients received decitabine (DAC) consolidation.With the median follow-up of 25.5 months (range, 1.1 to 33.5), the 2-year overall survival, leukemia-free survival (LFS) and cumulative incidence of relapse rates were 23.7%, 30.5% and 69.5%, respectively. Multivariate Cox regression analyses showed that a better LFS related to the absence of high-risk cytogenetics and genetic characteristics, DAC combination with fludarabine and cyclophosphamide lymphodepletion regimen, and bridging allo-HSCT or DAC consolidation.
Conclusions: Our study showed that the second infusion of tandem CD19/CD22 CAR-T therapy obtains a better response than other infusion strategies. Bridging allo-HSCT or DAC consolidation had a significant survival benefit in patients with CR after the second CAR-T therapy.
Evaluating the Impact of Prophylactic Intrathecal Chemotherapy in Diffuse Large B Cell Lymphoma (DLBCL) Patients with High Central Nervous System International Prognostic Index (CNS-IPI) Scores: A Comprehensive Review and Meta-Analysis
Authors: Al-Banaa Kadhim , Alshaikhli Alfarooq , Poudel Shyam , Shrotria Shiva , Wang Can , Hanona Paul , Gopishetty Swathi , Idogun Precious , Ezekwudo Daniel , Jaiyesimi Ishmael 
Published: December 2023

Introduction
Diffuse large B cell lymphoma (DLBCL) with a high CNS-IPI score poses an increased risk for relapse involving the central nervous system (CNS). Despite various studies seeking to validate the use of CNS prophylaxis, including intrathecal chemotherapy, results have been equivocal. This ambiguity may be partly attributed to the lack of prospective data, patient heterogeneity, limited sample sizes, and outdated methodologies in pre-rituximab studies. Consequently, administering CNS prophylaxis with intrathecal chemotherapy is not a standardized practice, and decisions are often based on individual factors such as anatomical location, disease stage, and specific case characteristics.
Methods
This systematic review and meta-analysis were conducted and reported using the PRISMA guidelines. A thorough search was conducted across multiple online databases, including Google Scholar, Science Direct, PubMed, Cochrane, CINAHL, EMBASE, ProQuest Dissertation and Theses, ISI web of knowledge, PsycINFO, and Embase for articles on this topic. The date range of the articles included was between 2013-2023 and were limited to English language, peer-reviewed articles, cohort studies, retrospective studies & randomized controlled trials. The inclusion and exclusion criteria for studies helped to determine suitable studies
Results
Out of 179 studies retrieved from the database search and 2 from other studies' reference lists, 6 met the inclusion criteria for qualitative and quantitative analysis. Data from 2209 patients in these 6 studies was analyzed using a random-effects model to perform a subgroup analysis of the benefits and risks of prophylactic intrathecal chemotherapy for patients with DLBCL and a high CNS-IPI score. The results demonstrated a significant improvement in overall treatment success, with a difference of 0.60 (95% Confidence Interval, 0.45 to 0.93) on a 0-10 Visual Analogue Scale (VAS). The heterogeneity of the studies was determined to be significant (P=0.02), further substantiating the positive impact of prophylactic intrathecal chemotherapy on the therapy success rate for DLBCL patients with high CNS-IPI scores.
Conclusion
Our review elucidates that intrathecal chemotherapy as a prophylactic measure can reduce the rate of CNS relapse and enhance overall survival for patients with DLBCL possessing a high CNS-IPI score. However, it underscores the necessity to thoroughly assess each patient's unique clinical situation when determining if intrathecal chemotherapy is the appropriate therapeutic approach.
VISTA a Potential New Immuno-Oncology Target in Acute Myeloid Leukemia
Authors: Guillaudeux Thierry , Sridhar Shaarwari , Frazier Emily , Ovechkina Yulia , Iadonato Shawn 
Published: December 2023

V-domain Immunoglobulin Suppressor of T cell Activation (VISTA/PD-1H) is a B7 family member highly expressed on circulating and intra-tumoral myeloid cells. VISTA is a negative checkpoint protein that inhibits anti-tumor T cell responses. In multiple solid tumors, VISTA expression is associated with poor overall survival and is also described as a potential mediator of resistance to anti-CLTA-4 and anti-PD1 therapies. VISTA expression was also investigated in hematological tumors and described as highly expressed on blasts including leukemic stem cells from AML patients. Therefore, VISTA is a unique and potentially new target for cancer immunotherapy in this indication.
Kineta has developed a fully human monoclonal antibody targeting VISTA, KVA12123, currently evaluated in a Phase 1/2 clinical trial alone and in combination with pembrolizumab in cancer patients with advanced solid tumors. We also believe that VISTA could be a promising target for AML patients. Therefore, we have tested in vivo, in a Kasumi-3 AML disseminated tumor model, the efficacy of our antibody alone or in combination with standard of care therapy, cytarabine and doxorubicin. Like leukemic cells in AML patients, the Kasumi-3 cell line exhibits a high level of VISTA expression on its cell surface. Female NOD-scid mice were first inoculated with Kasumi cells via tail vein injection, and tumor engraftment was evaluated at different time points in the blood, spleen and bone marrow to monitor engraftment using CD34, CD33, CD45 and HLA-DR cell surface expression markers. Then, when tumor burden was high, standard of care treatment (5 doses of 16mg/kg AraC by IV injections and 3 doses of 0.5mg/kg Dox by IV injections) was initiated in indicated groups for 5 cycles, before starting treatment with KVA12 for the rest of the study. Tumor engraftment was tested after chemotherapy alone to evaluate the efficacy before starting treatment with our antibody. Our results showed that untreated mice had higher number of Kasumi-3 cells in the blood when compared to chemotherapy treated mice. Two backbone versions of our antibody were then tested, a human IgG1 - KVA12123 and a human IgG4 - KVA12402, at 30mg/kg IP injections twice a week for 5 weeks. Our data showed that KVA12123 and KVA12402 as single agents and in combination with chemotherapy reduced Kasumi-3 tumor load in the blood, spleen and bone marrow within the groups and significantly improved survival when compared to IgG1 control groups.
These data all together indicate that anti-VISTA targeted therapy should be evaluated in myeloproliferative disorders like AML alone or in combination with standard of care.
In Vivo Generation of Functional CD19 CAR-T Cells Using a Serum-Stable CD3-Targeted Lentiviral Vector
Authors: Krotova Karina , Nenavath Gopal Naik , Packiriswamy Nandakumar , Vandergaast Rianna , Moy Melissa , Ziegler Christopher , Muñoz Alía Miguel A , Peng Kah-Whye , Russell Stephen 
Published: December 2023

The high cost and complexity of CAR-T therapy has spurred efforts to generate CAR-T cells directly within patients via systemic infusion of serum-stable lentiviral vector particles that target T cells in situ for CAR transgene delivery. To ensure effective in vivo delivery, these vectors need to show resistance to neutralization by human serum, demonstrate transduction specificity for T cells, and support subsequent expansion of CAR-T cells.
Lentiviral vectors (LV) pseudotyped with VSV G-glycoprotein (G-WT) transduce target cells via the LDL receptor (LDLR), which is ubiquitously expressed at variable levels and is upregulated on activated T cells. G-WT LVs are widely used to generate therapeutic CAR-T cells ex vivo, following purification and ex vivo activation of the T cells.
To generate LVs capable of targeting circulating T cells in vivo, we utilized rational engineering of the VSV-G glycoprotein to design a G-CD3. This modified G protein avoids LDLR recognition, incorporates a CD3-specific single-chain antibody for targeted recognition of T cells, and minimizes LV particle inactivation upon exposure to human serum. G-CD3 LVs were shown to target and transduce resting T cells in peripheral blood mononuclear cell (PBMC) cultures with very high efficiency. The G-CD3 LVs powerfully activated T cells, evident from their increased CD25 expression, and the infection was not hindered in the presence of fresh, complement-active human serum (25%). Additionally, G-CD3 LVs efficiently transduced T cells in fresh human whole blood without apparent off-target transduction of non-T cell lineages.
In vivo evaluation of G-CD3 LV efficiency was performed in humanized NSG mice by intravenous (IV) or intraperitoneal (IP) delivery of G-CD3 LVs encoding anti-CD19 CAR and GFP transgenes. Both IV and IP delivery of G-CD3 LV led to in vivo CAR T cell generation with peak levels of 17% of total circulating human CD45+ cells on day 21 post vector administration. The in vivo generated CD19-CAR-T cells were fully functional as evidenced by the disappearance of endogenous human CD19+ B cells from peripheral blood. Tissue analysis of spleen, bone marrow, and liver at the time of sacrifice (day 39 post LV injection), showed anti-CD19 CAR-T in all three tissues accompanied by an absence or dramatic reduction in tissue-resident human CD19+ B cells. In contrast, CAR-T cells were not detected at any time point in humanized NSG mice injected with G-WT LV, regardless of whether the virus was administered IV or IP.
In summary, these data highlight the potential applicability of fully retargeted, serum stable LVs for in vivo gene delivery or genome editing, offering promising prospects for future human gene therapy applications.
Spur Cell Anemia: A Rare and Under-Diagnosed Cause of Anemia in Alcoholic Patients with Advanced Liver Disease
Authors: Hajiabbasi Maryam 
Published: December 2023

Anemia in patients with alcohol use disorder (AUD) has a wide range of differential diagnoses, which can be challenging to interpret. It is often multifactorial, such as due to blood loss with or without coagulopathy, direct toxic effect of alcohol on hematopoietic cells, chronic inflammation, abnormalities in red blood cells (RBC) membranes, and nutritional deficiencies. Alcohol-induced liver disease may contribute to anemia through coagulopathy, splenomegaly, and hypersplenism. Macrocytosis is a common finding in patients with AUD due to B12 or folate deficiency or RBC membrane abnormality due to changes in lipid composition of RBC membranes. Patients with cirrhosis may have low haptoglobin due to impaired synthetic liver function. However, non-autoimmune hemolytic anemia and concomitant alcohol-induced liver damage should raise suspicion for spur cell anemia (SCA) and Zieve's syndrome. These medical conditions tend to be underreported or misdiagnosed.
We describe a 44 year old male patient with a past medical history of AUD who presented with alcohol intoxication and was found to have newly diagnosed decompensated cirrhosis with severe macrocytic anemia with Hgb of 5.9 g/dl. Patient denied overt bleeding. Initial workup revealed high absolute reticulocyte count of 10.8% with retic index of 3, low haptoglobin of &lt;15 mg/dl, elevated LDH of 275 u/l, negative Coombs test consistent with non-autoimmune hemolytic anemia, normal folate and B12 levels, and mixed direct and indirect hyperbilirubinemia. Peripheral blood smear showed anisopoikilocytosis, macrocytes, many spur cells, increased reticulocytes, but no schistocytes. Endo-colonoscopy showed grade 1 esophageal varices without evidence of recent bleeding. Given the history of heavy alcohol intake and cirrhosis, combined with Coombs-negative hemolytic anemia and peripheral blood smear findings of numerous spiculated red cells, our patient was diagnosed with SCA. At the time, due to active alcohol use, the patient was deemed not to be a candidate for liver transplant.
This case report aims to aid physicians with the recognition of acute hemolytic anemia in patients with alcoholic liver injury and consideration of SCA and Zieve's syndrome as potential underlying causes. Prompt recognition and differentiation between these medical conditions are crucial as prognosis and treatment are different. Early recognition of SCA and timely consideration of liver transplantation can significantly improve patient outcomes.
Characterizing Hereditary Hemolytic Anemias in a Hispanic Cohort Using Next Generation Sequencing
Authors: Chang Simone , Kasianchyk Mariia , Velazquez Vanessa , Borrego Andre , Maharaj Satish 
Published: December 2023

Background:
Hereditary hemolytic anemias (HHA) are a heterogeneous group of nonimmune disorders characterized by increased red blood cell (RBC) destruction due to abnormalities of hemoglobin stability, defects of RBC metabolism, and disorders of RBC hydration. Although uncommon, they comprise important causes of anemia in children and adults by causing chronic or episodic hemolysis. Clinical, laboratory, and genetic heterogeneity characterize this group of disorders and make recognition challenging. The advent of next generation sequencing (NGS) has made accurate genetic diagnoses attainable, with an increasing number of studies reporting clinical utility in HHA. For patients with suspected HHA, multigene testing is available incorporating an NGS panel associated genes. However, there is limited data on the use of multigene NGS to characterize HHA in the Hispanic population. Herein, we describe results from NGS testing of a pilot cohort referred for HHA investigation to hematology clinics along the US/Mexico border.
Methods:
Pediatric and adult hematology clinics at Texas Tech University in El Paso, TX collaborated for testing on HHA referral and clinical case review. The diagnostic panel (AnemiaID/PerkinElmer/Revvity®) included 51 genes encoding RBC cytoskeletal proteins, membrane transporter, RBC enzymes, and certain bilirubin metabolism genes. The complete coding region, splice site junctions, and, where appropriate, deep intronic or regulatory regions were covered. Targeted gene capture and library construction for NGS were performed using a Whole Blood and Saliva kit and 150 base pair paired-end sequencing was done on Illumina® NGS systems at target average coverage of 80×. NGS output data were summarized descriptively for all patients and clinically correlated for diagnosis.
Results:
In this pilot cohort, 11 patients were tested during September 2022 to June 2023, age ranging 7-75 years, with females accounting for 64% of the group. Findings are reported in the Table below; Pathogenic variants leading to a definitive diagnosis were identified in 8 of 11 cases (73%). The most frequently mutated genes were SPTA1 followed by PIEZO1, SPTB, RHAG, GSR, SLC4A1 and TPI1. Approximately 58% of the mutations were reported as novel. Diagnoses included, most commonly, Hereditary Spherocytosis followed by Hereditary Pyropoikilocytosis and Congenital Dyserythropoietic Anemia. Compound heterozygosity leading to complex interactions was noted in 4 cases (36%), involving SPTA1 low expression alleles in trans (alpha-LELY and alpha-LEPRA) in 2 cases.
Conclusion:
Hereditary hemolytic anemia in the Hispanic population is characterized by complex molecular interactions and a high prevalence of novel mutations in RBC cytoskeleton/enzyme genes. NGS has clinical utility and should be considered for accurate diagnosis. Further research into the molecular basis and genetic spectrum of HHA in the Hispanic population is needed.
Stage-Specific Erythropoietic Regulation By Long Non-Coding RNAs (lncRNAs) in Patients with Monge's Disease
Authors: Azad Priti , Zhou Dan , Rana Tariq M , Haddad Gabriel 
Published: December 2023

Background: Approximately 20% of individuals living in Andean mountains suffer from Chronic Mountain Sickness, CMS, or Monge's disease. They manifest the disease in adulthood and often die because of excessive erythropoiesis (EE) (Hematocrit of &gt;65%, averaging 72%, mostly occurring in males), an extreme phenotype that can often lead to myocardial infarction or stroke. The uniqueness of this Andean population is even more significant when we realize that there are individuals who live side by side at the same altitude as those with CMS but do not suffer from the same extreme phenotype (non-CMS).
Methods: To understand the mechanistic basis of EE in these patients, we performed RNA seq of cells from both CMS and non-CMS populations. We further utilized an in-vitro human iPS-derived model system to study the functional role of genes at various erythroid stages.
Results: Interestingly, we discovered a unique non-overlapping profile of differentially expressed long non-coding RNA (lncRNAs) between CMS and non-CMS cells. Among these lncRNAs, LINC00431 was significantly downregulated in non-CMS cells as compared to CMS in hypoxia (mimicking high altitude). We further validated the expression of LINC00431 and found it to be significantly downregulated under hypoxia in the non-CMS cells as determined by the qPCR (P&lt;0.01). We hypothesized that this lncRNA plays a role in regulating EE response at high altitude. Knock-down of LINC00431in the CMS cells did not affect the early stages of erythroid proliferation (such BFU-E) but there was a large effect in the later erythroid stages such as the CFU-E and reticulocyte stages (CD71+, CD235a+ cells). Particularly, KD of LINC00431 resulted in a major reduction (from ~60% to ~7%, p&lt;0.0001) of CD235a+ erythroid cells (Reticulocyte stage-a later erythroid stage). In contrast, we have also shown that the knock-down of LINC02228 has a large impact at the early erythropoietic stage BFU-E of the CMS cell reducing by 50% when we KD LINC00228.
Conclusion: These results show a stage-specific erythropoietic regulation by lncRNAs in the CMS and non-CMS patients. LINC02228 plays a critical role at the BFU stage and LINC00431 plays an important role particularly at later erythroid stages (CD71+, CD235+ Reticulocyte stage). This stage specificity also suggests distinct mechanisms for each lncRNAs and their downstream targets.
Automated Preselection of Regions of Interest By Deep Learning Facilitates Rapid Whole Slide Image Analysis of Bone Marrow Smears
Authors: Schneider Martin M. K. , Schulze Freya , Schmittmann Tim , Riechert Sebastian , Gediga Miriam Eva Helena , Sockel Katja , Kroschinsky Frank P. , Wendt Karsten , Bornhaeuser Martin , Middeke Jan Moritz , Eckardt Jan-Niklas 
Published: December 2023

The analysis of bone marrow smears (BMS) serves as a critical diagnostic tool in hematology, offering valuable insights into the cellular composition and distinctive morphology of bone marrow cells. It enables the accurate diagnosis, classification, and monitoring of various hematologic diseases. Currently, both clinical routine diagnostics and scientific research, especially for the implementation of digital technologies, rely heavily on the manual selection of regions of interest (ROI) suitable for cell-level evaluation. Current guidelines recommend the manual evaluation of up to 500 cells per sample making the process time-consuming, cost-ineffective, subjective, and prone to human error. Consequently, significant inter-observer variability and inconsistencies in diagnosis often arise.
Automated analysis of BMS using whole slide images (WSI) offers a promising approach to alleviate financial and time-related burdens while promoting standardization and streamlining of routine diagnostics. We have developed a machine learning approach utilizing a fully convolutional neural network (CNN) specifically designed for semantic segmentation based on the algorithm by Long et al. (IEEE Computer Society, 2015). The model is capable of learning deep semantic representations in its classification backbone combined with shape features in a shallow layer to produce detailed segmentations. The classification backbone consists of an adapted residual neural network with 50 layers (Resnet50) by He et al. (arXiv:1512.03385, 2015). To train the model, we curated a diverse dataset consisting of 60 different WSI BMS, each manually and independently annotated by at least two experts providing a robust ground truth (figure 1). Data augmentation was performed via horizontal flipping, random resizing, and random cropping. A train-test split of 80:20 was implemented with 5-fold internal cross-validation. The model's accuracy was evaluated as mean intersection-over-union ratio yielding high congruency between ground truth and model prediction (figure 2). The final model uses downscaled images effectively reducing data storage size and data transfer time decreasing fully automated ROI prediction time to &lt;300 ms for one WSI.
The model allows for in-depth examination at the cellular level, harnessing the potential of thousands of relevant cells present in a WSI, in contrast to the limited range of 200 to 500 cells typically examined in conventional clinical routine diagnostics. Thereby, the model solves an essential bottleneck in computer vision in microscopy minimizing human manual labor and standardizing BMS processing results.