bluuebunny/crossref_metadata_2025 huggingface dataset:
A Pragmatic Approach to Navigating FDA Guidance for Therapeutic Product Development in Malignant Hematological Disease
Authors: Wang Limin , Johnson Angela N
Published: December 2023

Background: Owing to the unique therapeutic characteristics and diverse etiology of underlying disease, navigating available regulatory guidance can be a challenge for newcomers to therapeutic research and development (R&D). Further, classes of drugs and biologic products are regulated separately by the U.S. Food and Drug Administration (FDA) centers CBER and CDER, respectively.

Methods: We provide background on the FDA guidance structure herein and further examine publicly registered clincialtrials.gov data to provide an overview of development status by phases and novel modality (i.e., CAR-T) in context of the regulatory landscape.

Results: Therapeutic products developed for malignant hematological disease can be regulated by either CDER or CBER depending on the type of drugs, ranging from small molecules to macromolecules, including cellular and gene therapies. In 2019, CDER of Hematology and Oncology Products (OHOP) has been reorganized and renamed the Office of Oncologic Diseases (OOD). The new OOD structure consists of six divisions including Division of Hematologic Malignancies 1 (DHM1) and Division of Hematologic Malignancies 2 (DHM2) for the review of hematologic malignancies. In 2022, CBER of Office of Tissues and Advanced Therapies (OTAT) was transformed into the Office of Therapeutic Products (OTP) to address the exponential growth in cell and gene therapies. PDUFA VII expanded the INTERACT meeting program to include both centers, a major change that may drive early phase development collaboration.

A review on the current state of hematologic malignancies clinical trials in USA shows that, of 301 hematologic malignancies related trails in USA registered to ClinicalTrials.gov (Fig. 1), 4 are in early phase 1(exploratory phase), 126 in phase 1, 120 in phase 2, 18 in phase 3, and 1 in phase 4. Most of the trials are still in early phases 1 or 2. Among the 301 registered clinical trials, 21 of clinical trials (Fig. 2) are CAR-T related, and 10 of the trials are gene modified cell products related.

Conclusions: Compared with traditional treatment on malignant hematological disease, immunotherapy is the upcoming trend in cancer treatment. Chimeric antigen receptor (CAR) therapy is at the forefront of cellular/adoptive immunotherapy for cancer treatment due to its sustained remission and better quality of life. Much research and clinical trials have been undertaken in recent years which led to several successful outcomes in CAR therapy. Despite tremendous progress, CAR-T are still faced with many challenges, including evolving regulatory framework along the road. We summarize the regulatory landscape to assist therapy developers to align with FDA expectations.
A Retrospective Analysis on the Efficacy and Saftey of Obinutuzumab in the Treatment of Non-Hodgkin B Cell Lymphoma
Authors: Zhao Yangzhi , Guo Wei , Li Jia , Wang Xingtong , Wan Xin , Li Junna , Bai Ou
Published: December 2023

Backgroud: Obinutuzumab was approved by China's Drug Administration in June 2021, and there have been few real-world reports about efficacy and safety.

Objective: Analysis the efficacy and safety of Obinutuzumab in the treatment of non-Hodgkin B cell lymphoma in our center.

Methods ： We analysed the non-Hodgkin B cell lymphoma patients who received Obinutuzumab combined with chemotherapy or maintenance treatment during September 2021 and July 2023, median follow up 9 months（1-20）, to evaluate efficacy and safety.

Results: In all 165 patients, male 88（53%), female 77（47%）, median 56 (26-83) years. FL 80 (48%), MZL 22 (13%), CLL/SLL 13 (8%), DLBCL 42 (25%), others 8 (5%). Newly diagnosed patients 114 (69%), relapsed/refractory 51 (31%). 124 patients received Obinutuzumab combined therapy, and 42 (25%) patients received Obinutuzumab maintenance treatment. Median treatment 4 cycles（1-11）. I/II 33 (20 %), III/IV 131 (79 %). Low-medium group 91（55%）, medium-high group 74（45%）. ORR 66.7%, CRR 40.6%. 3 patients died of progression of disease. 1 elder patient died of pneumonia. Further subgroup analysis revealed, ORR（78% vs 22%, p<0.001）and CRR（81% vs 19%, p=0.02）of newly diagnosed patients were better than relapsed/refractory patients. There was no difference in CRR among different stages patients（I/II vs III/IV, 24% vs 76%, p=0.424）, ORR（58% vs 42%, p=0.373）and CRR (63% vs 37%, p=0.481）between low-medium and medium-high patients. The patients received more than 4 cycles treatment with better ORR and CRR. The intensive treatment of Obinutuzumab in first cycle could improve ORR (87% vs 13%, p=0.036,）and decreased IgM ( p=0.0016）, but didn't not affect CRR （87% vs13%, p=0.343）, IgG ( p=0.13） and IgA ( p=0.41）. Maintenance treatment decreased IgM ( p=0.00064）but not for IgG or IgA. In treatment related side effects, any adverse events were 42（25%）, more than grade III were 18（11%）, tumor lysis syndrome 11 (7%), only one patient need blood purification treatment and recovered. Liver injury 8 (5%) . All the infusion-related response 23（14%）were grade I-II. Only 2 patients delayed treatment because of the side effects.

Conclusion: Obinutuzumab is safe and effective in the treatment of non-Hodgkin B cell lymphoma, even for the III/IV or medium-high group patients will have similar results with I/II or low-medium patients. Further follow-up is still needed to determine the long-time efficacy and safety.

Key words: retrospective analysis,Obinutuzumab, non-Hodgkin lymphoma.
Efficacy and Safety of Polatuzumab Vedotin in Follicular Lymphoma: A Systematic Review of Clinical Trials
Authors: Jumean Samer , Aiman Wajeeha , Qirem Murad , Alkhlaifat Osama , Ali Muhammad Ashar , Shaaban Hamid , Guron Gunwant , Al Omour Bader , Ramahi Amr , Isbeih Nooredeen
Published: December 2023

Introduction:

Polatuzumab vedotin (Pola) is a humanized anti-CD79b monoclonal antibody linked with antimitotic agent monomethyl auristatin E. On internalization, the linker is cleaved, and antimitotic activity inhibits the cell division and growth of cancer cells. Pola is tested in multiple combinations in relapsed/refractory (RR) follicular lymphoma (FL) in recent years. In this systematic study, we will assess the efficacy and safety of Pola based combinations in RR FL.

Methods:

A literature search was performed on PubMed with keywords, “Polatuzumab vendotin” and “lymphoma” from the inception of data till 07/01/2023. PRISMA guidelines were followed while conducting this systematic review. We reviewed 160 articles and included 1 randomized clinical trial (RCT, N=41), and 4 non-randomized clinical trials (nRCT, N=163) based on inclusion criteria. Data was extracted for complete response (CR), partial response (PR), overall response (ORR), progression free survival (PFS), overall survival (OS) and ≥grade 3 side effects.

Results:

In 5 trials (N=204), 13 patients with Pola + atezolizumab (Ate) + obinutuzumab (Obi), 49 patients with Pola + Obi + venetoclax (Ven), 39 patients with Pola + Obi, 62 with Pola + Obi + lenalidomide (Len), and 20 patients with Pola + rituximab (Rit). In an RCT by Morschhauser et al. (N=41), Pola + Rit was compared with pinatuzumab vedotin (Pina) + Rit. ORR, CR, PR, and mPFS was 70%, 45%, 25%, and 15.3 months, respectively with Pola + Rit, as compared to 62%, 5%, 57%, and 12.7 months, respectively, with Pina + Rit. In a trial by Philips et al. (N=39), ORR, CR, PR, and mPFS were 67%, 36%, 31%, and 11.5 months, respectively. In 3 nRCTs (N=124), three drug regimens were used with Pola and Obi as backbone. ORR, CR, PR were 50%-83%, 20%-61%, and 20%-39%. Table 1. ≥ Grade 3 hematological and non-hematological side effects are given in table 1.

Conclusion:

Pola was well tolerated by most of the patients with FL alone or in combination with Obi, Rit, Len, Ven. Pola had relatively better response and survival rates as compared to Pina in combination with Rit. In early phase clinical trials, three drug regimens of Pola and Obi with Ven, and Len were effective in the treatment of heavily pretreated follicular lymphoma. In an early phase trial, Pola + Obi + Ate had limited efficacy in FL with safety concerns. More randomized double blind multicenter phase III clinical trials are needed to confirm these results.
This Is a Title in Title Case: A Retrospective Study for 14 R/R DLBCL Patients
Authors: Shen Jianzhen , Li Xiaofan
Published: December 2023

This study retrospectively analyzed 14 R/R DLBCL patients who received combined treatment of obutenib in Fujian Medical University Union Hospital from July 2021 to April 2023. Except for four patients using Orelabrutinib as post-second line treatment, all the other patients received 1 to 8 course Orelabrutinib as second line treatment. The combined chemotherapy regiments include R-EPOCH and R-DHAP. Among them, there were 8 males (57.1%) and 6 females (42.9%), with a median age of 56/57 (39-79) years. The median follow-up time was 11 (3-27) months. At the time of diagnosis, there were 12 cases (85.7%) in the Ann Arbor stage III-IV, 7 cases (50.0%) with an IPI score of 3-5, 6 cases (42.9%) with non GCB type, 8 cases (57.1%) with double expression, 2 cases (14.3%) with primary central nervous system lymphoma, and 6 cases (42.9%) with secondary central nervous system lymphoma. All patients had extranodal involvement, with 10 cases (71.4%) with ≥ 2 extranodal lesions and 8 cases (57.1%) with elevated LDH. At the end of treatment, the ORR and DCR were 42.9% and 28.6%, respectively, with a median PFS of 7 (95% Cl: 4-12) months and a median OS not achieved. Among them, the ORR and DCR of central nervous system lymphoma were 75.0% and 50.0%, respectively, with a median PFS of 9.5 (95% Cl: 6-22) months, and the median OS was not reached. Neutropenia (n=2, 14.3%) is the most common level 3/4 adverse event. No patient died due to adverse reactions.This study suggests that the combination of Orelabrutinib and chemotherapy has an excellent therapeutic effect in R/R DLBCL, especially in CNSL.
Real-World Treatment Patterns, Discontinuation and Clinical Outcomes in Patients with B-Cell Lymphoproliferative Diseases Treated with BTK Inhibitors in China
Authors: Yan Yuting , Lyv Rui , Wang Tingyu , Yu Ying , Huang Yanshan , Xiong Wenjie , An Gang , Zou Dehui , Qiu Lugui , Yi Shuhua
Published: December 2023

Bruton tyrosine kinase inhibitor (BTKi) has demonstrated substantial efficacy in treating B-cell lymphoproliferative diseases (BLPD). Nonetheless, the significant discontinuation rates due to toxicity or financial reasons cannot be overlooked. In China, empirical evidence on the usage of BTKi remains scarce. To address this, a retrospective cohort study was conducted focused on 673 Chinese patients with BLPD who underwent at least one month of BTKi therapy. Median age at BTKi initiation was 60 years. The median duration on BTKi treatment of the whole cohort was 36.4 months. The median post-BTK survival was not reach. BTKi-based treatment was permanently discontinued in 288 (43.8%) patients during follow-up, mostly attributed to progressive disease. Within the first 6 months of BTKi treatment, 76 patients (26.3%) had early treatment discontinuation. Patients with early discontinuation had extreme worse outcome with a median post-discontinuation survival of only 6.9 months. On multivariate analysis, withdrawal BTKi by toxicity and withdrawal BTKi within 6 months retained to be independent predictors of post-BTK survival, after taking account of the response depth, lines of therapy and baseline cytogenetics including 17p deletion. The decision between BTKi monotherapy and combination therapy, along with the preference for first or second-generation BTKi, exerted no significant impact on survival. These observations contribute valuable real-world insights into the utilization of BTKi in China. We concluded that BTKi is an effective and well-tolerated treatment for long-term use in Chinese patient population. However, it is imperative to stress that a proportion of patients discontinue BTKi early, leading to suboptimal outcomes. This study underscores the importance of adherence to BTKi therapy for improved clinical outcomes in real-world patients.
Interplay between Spatial Confinement of Thrombin Activity and Dense Granule Secretion Regulates Dynamics of Thrombus Formation
Authors: Masaltseva Anastasiia , Panteleev Mikhail A. , Nechipurenko Dmitry
Published: December 2023

Background

Thrombi formed in healthy murine microvessels have a heterogeneous structure, comprising a thrombin-dependent core and a shell supplied by platelet-derived agonists, such as ADP from dense granules. After the injury is inflicted, the core steadily grows until it reaches a plateau in size, while the shell follows three stage dynamics of rapid growth, shrinkage and stabilization. However, it is not clear how the complex dynamics of the overall thrombus structure is orchestrated by various cues.

Aim

We aimed to analyze whether spatiotemporal kinetics of thrombin-induced dense granule secretion is responsible for the observed dynamics of thrombus formation in microvessels.

Methods

We developed a multiscale computational model of thrombogenesis in response to a vessel wall injury. Platelets were modeled as two-dimensional particles, while platelet agonists were described as virtual particles, which induced platelet activation through affecting mechanical interactions between platelets and triggering platelet degranulation. As a platelet aggregate grew, the blood flow was recalculated. The model was built in a bottom-up manner and its description of single-cell processes was based on the experimental findings.

Results

The model showed that thrombin produced at the injury site was mostly confined to the innermost layers of the thrombus due to competing intrathrombus diffusion and flow-mediated dilution. Such spatial confinement of thombin resulted in the formation of a thrombus with a pronounced thrombin-activated core where platelets were able to secrete their dense granules. Released ADP recruited new platelets and triggered the formation of a thrombin-free thrombus shell. The thrombus grew until it ruptured under the hydrodynamic force and the growth repeated in cycles. However, our results predicted that if the propagation of thrombin along the flow was limited either by volumetric or surface inhibition, the cycles of thrombus formation stopped once the dense granule pool in the core was depleted. Without the supply of newly secreted ADP, the thrombus shell disassembled and the thrombus size was stabilized, in line with experimental findings.

Conclusion

Computational modelling predicts that thrombin-induced dense granule secretion is responsible for the rapid formation of the thrombus shell, which disintegrates once the granule pool is depleted due to intense secretion in the confined thrombus core. We suggest that this mechanism is responsible for the three-stage dynamics of thrombus growth in vivo.
Title: Single-Cell Transcriptome Analyses Further Elucidate the Mechanism of Action of Eed Inhibition on HbF Induction
Authors: Han Summer Xia , Fitz Emily M , Allen Allysa , Choudhuri Avik , Sartain Caroline V , Bruno Paul A , Jacobs Jeffrey W , Stuart Billy
Published: December 2023

Mutations in the gene encoding the hemoglobin β (HBβ) subunit lead to red blood cell disorders such as Sickle Cell Disease (SCD) and β-thalassemia. Hemoglobin γ subunit, a fetal counterpart of HBβ, can relieve the disease symptoms by forming a functional complex with hemoglobin a subunit, thereby compensating for the mutated HBβ. The expression of β-like hemoglobins is tightly regulated during development, with a switch from the fetal ortholog (γ) to the adult ortholog (β) shortly after birth. Maintenance of HBγ is therapeutically beneficial for SCD patients and is observed clinically in individuals with mutations for both SCD and hereditary persistent fetal hemoglobin (HPFH) who exhibit less severe or even no SCD-related symptoms. Polycomb repressive complex (PRC) proteins have been shown to regulate the expression of β-like hemoglobins. Our previous research has demonstrated inhibitors targeting the PRC2 protein embryonic ectoderm development (EED) upregulate fetal hemoglobin (HbF) expression, leading to the formation of HbF complexes in both human CD34 + HSC-derived erythroid cells and the Townes SCD mouse model. However, the functional mechanisms of PRC complexes in this regulatory process and their target genes remain to be clarified.

In our study, we conducted single-cell transcriptome analyses on erythroid cells derived from CD34 + HSCs collected at various stages of differentiation. We identified several erythroid subpopulations, from early progenitors to maturing cells, within the CD34 + derived erythroid cell cultures. HbF + and HbF - cell populations were also identified using known markers.

To explore the mechanisms underlying HbF upregulation by EED inhibition, we compared single-cell transcriptome profiles of samples treated with and without an EED inhibitor. Our findings reveal that EED inhibitor treatment consistently leads to increased HbF + populations across multiple donors without impacting erythroid cell differentiation.

Differential gene expression analysis revealed sequential changes in the expression of previously identified HbF regulators during differentiation in the presence of EED inhibitor treatment. Through pseudotime analyses, we identified erythroid progenitor cells undergoing fate bifurcation, leading to an increase in the lineage of mature cells that express HBγ. Finally, by comparing the pseudotime trajectories of different samples, we identified genes whose expression changed along the differentiation trajectories in response to EED inhibitor treatment which influence cell fate. Overall, this study provides a deeper understanding of the relevant biological pathways that respond to EED inhibition and potential for the identification of novel molecular targets for HbF induction.
Has the Survival in Patients with Multiple Myeloma Changed with the Advent of Latest Treatments?
Authors: Desai Krishna , Iqbal Sabah , Rather Manzoor , Thirumaran Rajesh
Published: December 2023

Introduction:

Multiple Myeloma (MM) is a hematologic malignancy of plasma cells. Over the last couple of decades, there have been groundbreaking advances in its treatment, emergence of chimeric antigen receptor T-cell (CAR-T) therapies and bispecific antibodies (bsAbs). As per National Comprehensive Cancer Network's (NCCN) current guidelines, the current standard of treatment is a 3-drug regimen bortezomib/lenalidomide/dexamethasone. The year 2003, marked the beginning of the revolution in the MM biological treatment. The aim of this retrospective study is to analyze the trends of multiple myeloma, particularly to assess the difference in survival, with the innovation in its novel treatment options.

Methods:

The data was collected from Surveillance, Epidemiology and End Result database Research. We extracted Multiple Myeloma cases diagnosed between 01-85+ years of age, using Incidence- SEER Research Date 17 Registries, Nov 2022 Sub (2000-2020), with ICD 0-3 Hist/behavior, malignant, code 9732/3 for plasma cell myeloma. The analysis was stratified based on age, sex, race, & survival curves based were compared, using the Log-rank test (GraphPad Prism).

Results:

A total of 109222 cases of multiple myeloma were identified between 2000-2020. Of these, ~55.18% were males and ~44.81% were females. Median age at the time of diagnosis is 69 years. The incidence rate overall has been increasing and was noted to be 62.38 per 1,000,000 population vs 56.38 in the year 2000. When stratified further on gender, incidence rates for males were noted to be up-trending from 69.32 in 2000 to 75.87 in the year 2020. Whereas for females the incidence rates in females in 2000 was at 47.02 and at 51.51 in 2020. The survival was compared for the general population before and after the emergence of newer treatment in the year 2003. There was a statistical difference found in the median of survival (mOS) before the introduction of the biologicals in 2003, was 25 months, and after the year 2003, was 46 months (P Value <0.0001, Hazard Ratio 1.451 95% Confidence Interval 1.421 to 1.481).

Conclusion:

Multiple Myeloma is a relatively rare malignancy, with a slightly stronger male predilection. Its overall incidence has been noted to be up-trending within the last 20 years, in the United States. The median age for diagnosis is 69 years. In the year 2003, FDA approved Bortezomib for the treatment of MM, which primarily targeted the proteasome and the ubiquitin proteasome system (UPS). This established the role of UPS in MM, and further led to the invention of newer biological treatment options for MM. When survival months were compared in the pre-and-post biological treatment era, it was noted that the mOS was only 25 months, whereas a significant increase in mOS to 46 months was noted, after the introduction of biological therapy in 2003. With newer groundbreaking research and approval of CAR-T therapies, we are on our way to making MM, a curable malignancy.
HbF Inducing Eed Inhibitor Ftx-6058 Selectively and Reversibly Impacts Bone Marrow in Wild-Type Mice
Authors: Fitz Emily M , Allen Allysa , Edwards Lynn , Raghunathan Suchi , Steward Briana , Matson David J , Sartain Caroline V , Bruno Paul A , Jacobs Jeffrey W , Stuart Billy
Published: December 2023

Sickle cell disease (SCD) is a genetic disorder characterized by red blood cell sickling, hemolysis, vaso-occlusive crises (VOCs), and other complications resulting from a mutation in the HBB gene. Increasing fetal hemoglobin (HbF) levels has shown promise in mitigating disease-related pathophysiology. In individuals with SCD who exhibit hereditary persistence of fetal hemoglobin (HPFH) and have HbF levels ranging from 20% to 30%, a remarkable reduction in symptoms is observed, highlighting the protective effect of increased HbF in SCD.

Preclinical and clinical findings indicate that compounds inhibiting the embryonic ectoderm development (EED) component of polycomb repressive complex 2 (PRC2) leads to elevations in HbF to therapeutic levels. These results suggest the potential for a substantial benefit for individuals living with SCD. To further understand the effects of PRC2 inhibition, this study aims to characterize the onset and reversibility of EED inhibition by FTX-6058.

Results from in vivo studies in wild-type mice demonstrated upregulation of the mouse fetal hemoglobin ortholog mRNA (Hbb-bh1) as well as target engagement in bone marrow though quantification of H3K27 trimethylation during seven days of active dosing of FTX-6058. Target engagement and differential gene expression profiles in the bone marrow subsequently returned to baseline shortly after the dosing period. These findings demonstrate that inhibition of EED through FTX-6058 leads to a minimal, transient impact on the bone marrow in mice, and any transcriptional changes are rapidly reversible following cessation of dosing. A small molecule EED inhibitor has the potential to provide a new treatment for sickle cell disease without irreversibly altering the bone marrow.
Prevalence of Sleep Disordered Breathing in Adult Sickle Cell Clinic Patients and Cardiovascular Correlates
Authors: Whitesell Peter , Mehari Alem , Mellman Thomas , Stokes Daniel , Mirzada Freba , Rock Angela , Taylor James
Published: December 2023

Sleep disordered breathing (SDB) is strongly correlated with hypertension and cardiovascular disease in the general population, and especially in African Americans. In Sickle Cell Disease (SCD), the adverse effect of nocturnal hypoxemia is anticipated to have additional adverse effects with the sickling of red blood cells and microvascular thrombosis unique to the disease. Prospective data regarding the prevalence and consequences of SDB in this population are lacking.

In an ongoing study, 27 adult subjects (age > 18 years), with homozygous HbSS genotype, presenting to the Howard University Sickle Cell Center for routine follow up have been recruited irrespective of the presence of sleep symptoms. Exclusion criteria include previous diagnosis or treatment of a sleep disorder or requirement for supplemental oxygen. Testing occurs when subjects are in a stable phase of disease, with no hospitalizations or medication changes for 4 weeks. In addition to clinical history and questionnaires, participants are assessed with blood work, 6 minute walk testing, pain and sleep diaries during 1 week actigraphy, transthoracic echocardiography and overnight in lab polysomnography with capnography.

At the time of this interim analysis, the mean age is 42 (range 29-71, +/- std dev 11) and male:female distribution is 17:10. The mean BMI is 20.4 (+/- 4.5). Mean Hgb is 9.1 g/dl (+/- 1.8) and mean LDH is 325 iu/L (+/- 163).

Polysomnography scoring utilized a 3% desaturation criterion for hyponeas as per American Academy of Sleep Medicine Scoring Manual Chapter VIII, 1A. Mean sleep efficiency is 77% ( std dev +/- 4.2%). 48% of subjects demonstrate some degree of sleep disordered breathing as defined by an AHI > 5 events/hour (33% mild, 4% moderate, 11% severe). 70% of subjects demonstrate an RDI > 5 events/hour (40% mild, 19% moderate, 11% severe). Twelve participants (44%) exhibit more than 5 minutes sleep duration at saturations <89% (mean = 187 minutes). Hypercapnia is not suggested as an important component of SDB with the mean ET CO2 = 40.7 mm Hg (range 30-46 mm Hg).

Echocardiographic values (LVPWd, Lad, MV E/A, PVV, and RVSP) do not correlate with any indices of sleep disordered breathing other than minutes <89% saturation correlating with RVSP.

In summary, this interim analysis of sleep in a population of unselected Sickle Cell Clinic patients demonstrates a striking frequency of sleep disordered breathing. This is despite a generally low BMI for these patients which is typically considered one of the most important risk factor for sleep apnea. Ongoing recruitment is planned to confirm these findings. Correlation with clinical outcomes and potential response to therapeutic interventions will be important future research questions.
Development of Organoid-Based Cytotherapeutics for Taste Loss Induced By Anti-Cancer Chemotherapy and Radiotherapy
Authors: Kim Donghoon , Park Sung-Soo , Jeong Yong-Taek , Kim Jin-Woo , Kim Heeje
Published: December 2023

Introduction

Ulcerative mucositis and taste loss are common side effects induced by cancer therapy, including chemotherapy and radiation therapy. These conditions cause significant oral pain and loss of taste, impacting food intake and physical health abnormalities such as malnutrition, as well as depression and mental health deterioration, which can affect prognosis. To address these challenges, an appropriate treatment for oral mucositis is required. While various types of oral mucositis treatments have been developed (e.g., patch, spray, ointment), most show indirect healing effects such as pain control and inflammation relief. Thus, a direct method for treating oral lesions should be developed. This research was supported by the Korean Fund for Regenerative Medi- cine (KFRM) grant funded by the Korea Government (the Ministry of Science and ICT, the Ministry of Health & Welfare) (21C0712L1-11).

Methods

To compensate for the difficulty in supplying human tissue, we conducted preliminary experiments using mice to establish protocols for manufacturing human oral tissue-derived cell therapeutics. we devised an organoid-derived cell sheet transplantation. First, we generated organoids derived from oral mucosa and prepared a cell sheet using a temperature-responsive cell culture dish. The harvested cell sheet was transplanted to the lesion site to enable structural and functional recovery of the oral mucosa and gustatory organs.

Results

We have developed a cell-therapeutic platform based on regenerative medicine technology to recover oral lesions in cancer chemotherapy and radiotherapy patients. Specifically, we developed a method for producing autologous cell therapeutics. The oral mucosa tissue of a patient scheduled for anticancer treatment was collected, and epithelial cells were obtained from the tissue. Using these cells, we generated organoids with differentiated taste bud cells, and isolated cells from the organoids were seeded onto temperature-responsive cell culture dishes and cultured beyond confluence for the formation of a sheet. We performed immunostaining to detect various types of taste cells in the cell sheet. We also confirmed the success of cell sheet transplantation and the subsequent healing effect.

Conclusion

Improving the prognosis and acquiring the will to recover of cancer chemotherapy and radiotherapy patients through repairing taste function and rapid healing of lesions are crucial. We have developed a cell therapy technology that enables the structural and functional recovery of the oral mucosa and gustatory organs, providing a promising solution to treat the side effects of anti-cancer treatment. Our research provides an R&D initiative for the treatment and prevention of intractable diseases by culturing, manufacturing, and transplanting living autologous cells.
Engineering Off-the-Shelf Gamma Delta CAR T Cells for the Treatment of Acute Myeloid Leukemia
Authors: Dwivedi Alka , Fu Lynn , Chien Chris Daniel , Pouzolles Marie , Shah Nirali N. , Taylor Naomi
Published: December 2023

Chimeric antigen receptor T cell therapy (CART) therapy has shown remarkable success in the treatment of B cell acute lymphoblastic leukemias (B-ALL) and lymphomas. However, CART therapies for acute myeloid leukemia (AML), where 5-year survival rates are significantly lower than for B-ALL, are only in their infancy. CD33-CART have potent activity against AML in preclinical models and a first-in-child/first-in-human phase 1/2 CD33-CART clinical trial for AML is ongoing in the Pediatric Oncology Branch of the National Cancer Institute (NCT03971799). Nonetheless, published outcomes suggest a modest efficacy of approximately 50% (Shahzad et al., Front Immunol 2023), highlighting the critical need to develop new strategies to improve CART accessibility and a more robust anti-AML response. We hypothesized that off-the-shelf gamma delta (γδ) CD33 CART cells could potentially overcome current challenges for the treatment of AML. γδ lineage T cells are unconventional lymphocytes whose functions are not restricted to MHC-mediated antigen presentation; they are primed for immediate responses, including tumor killing. Furthermore, allogeneicγδ T cells have the potential to induce robust anti-tumor cytotoxicity without causing graft versus host disease (GVHD).

Here, we generated γδ CAR T cells from healthy donor elutriated lymphocytes by activation with zoledronic acid and IL-2 for 7-14 days. Within 9 days post stimulation, the vast majority of lymphocytes were Vδ2+ and 30-40% were successfully transduced with a lentiviral CD33 CAR construct harboring the 4-1BB costimulatory domain. Importantly, and unlike conventional alpha beta (ab) T lymphocytes, >98% of these γδ CD33CAR T cells expressed IFNγ under basal conditions. This characteristic likely accounted for the efficient in vitro killing of AML cell lines by untransduced γδ T lymphocytes under conditions of high effector/target (E/T) ratios. While untransduced γδ T cells did not exhibit cytotoxicity following repeat AML stimulations, γδ CD33CAR T lymphocytes exhibited proficient in vitro cytotoxicity, with killing rates that were more rapid than those initiated by ab CD33 CART (Figure 1). These characteristics were associated with a prolonged metabolic activity of γδT cells; γδ CD33 CART expressed high levels of the GLUT1 glucose transporter for >14 days post activation whereas GLUT1 levels on ab CD33 CART returned to resting within 10 days. High GLUT1 levels were linked to efficient killing under conditions of basal glucose levels. Most notably, γδ CD33CAR T lymphocytes achieved high in vivo cytotoxicity, assessed using bioluminescent AML cell line xenografts in humanized NSG mice. Together, these data highlight the feasibility of generating allogeneic γδ CD33CART with a strong anti-AML cytotoxic response.
Investigating the Vasodilatory Effects of Tgpolyhb: A Potential Hemoglobin-Based Oxygen Carrier for Sickle Cell Disease and Thalassemia
Authors: Munoz Carlos J , Lucas Daniela , Muller Cynthia R , Gopal Srila , Cabrales Pedro
Published: December 2023

Background: Hemoglobin-based oxygen carriers (HBOC) hold promise in improving oxygen delivery to organs and tissues, potentially reducing the need for transfusions and associated complications, thus presenting an alternative approach to managing anemia.

Methods: We created a polymerized human hemoglobin called TgPolyHb, using transglutaminase, which has demonstrated effective oxygen transport and blood flow maintenance in animal studies. Our hypothesis suggests that TgPolyHb will not induce vasoconstriction or hypertension, side effects observed in previous generations of HBOC, such as HBOC-201. In the present study, we assessed the potential of TgPolyHb to reduce the side effects previously seen with HBOC-201, comparing it to stroma-free acellular hemoglobin (hHb - negative control) in Golden Syrian Hamsters. The animals were instrumented with a metallic frame with a small cover slip placed over the dorsal skin fold to observe and quantify microcirculatory health following a toapload infusion (arterial injection of 10% of their blood volume).

Results: The innovative HBOC, TgPolyHb, exhibited vasodilation effects in comparison to HBOC-201 and hHb, both exhibited a 10% reduction in diameter compared to baseline. Furthermore, TgPolyHb demonstrated a 10% increase in functional capillaries compared to HBOC-201 and hHb. Functional capillary density is an important marker of tissue health.

Conclusions: Our study provides preliminary data that could lead future investigations of HBOC such as TgPolyHb in decreasing transfusion burden in severe anemias.
Implementation of 4T Score Documentation on PF4 Testing in Patients with Suspected Heparin-Induced Thrombocytopenia
Authors: Ruiz Maria Andreina , Gopalani Radhan , Justiz Jessica , Clarke Heidi , Dittmar Erika
Published: December 2023

Purpose: Heparin-induced thrombocytopenia (HIT) is a severe complication of heparin therapy. Diagnosis requires clinical evaluation and laboratory testing. The 4T score is a pretest clinical scoring tool used to determine the probability of HIT. A non-profit health system implemented mandatory documentation of 4T score calculation prior to ordering PF4 tests for suspected HIT patients. The purpose of this performance improvement project is to evaluate the implementation of 4T score documentation on PF4 testing and argatroban utilization.

Methods: This was a retrospective, multi-site chart review of patients conducted to assess the impact of mandatory 4T score documentation prior to a PF4 order. The study included adult patients who were admitted to one of the practice sites between January 2020 and November 2022 with a suspicion of HIT and a PF4 order. The primary outcomes included the number of PF4 tests ordered and the percentage of positive PF4 results. Secondary outcomes included number of SRA orders and percentage of positive SRA results, turnaround time for SRA, and the utilization of argatroban. Data collection included baseline demographics, documentation of 4T score, PF4 results, SRA results, documentation of heparin allergy, initial anticoagulant prescribed, platelet trend, ordering provider, and duration of argatroban therapy in patients with a positive PF4 result. The analysis involved using χ2 test and Fisher's exact test for the primary outcomes.

Results: Mandatory 4T score documentation significantly reduced PF4 orders by 68% over a two-year period (p<0.0001). It also improved the percentage of positive PF4 results from 12% to 24% (p=0.60) and positive SRA results from 19% to 44% (p=0.35). Notably, after documentation of the mandatory 4T score tool, only 21% of the patients had a PF4 order placed. A total of 42 patients with a positive PF4 test were included to assess argatroban utilization. The turnaround time for SRA in these patients was 3.6 days while duration of argatroban therapy was 6.8 days.

Conclusion: Implementation of mandatory 4T score documentation significantly reduced PF4 orders and improved HIT diagnostic testing in a non-profit health system. Challenges remain with SRA turnaround time and corresponding duration of argatroban therapy. The results of this project have demonstrated that standardized documentation is essential for optimizing HIT diagnosis and management protocols. Future efforts will focus on optimizing confirmatory HIT testing and non-heparin anticoagulation therapy.
CD70 CAR NK Cells in the Treatment of Multiple Myeloma
Authors: Lin Paul , Reyes Silva Francia C. , Lin Pei , Gilbert April L. , Acharya Sunil , Nunez Cortes Ana K. , Banerjee Pinaki , Fang Dexing , Melo Garcia Luciana , Daher, MD May , Basar Rafet , Patel Krina K. , Lee Hans C. , Orlowski Robert Z. , Rafei Hind , Marin David , Champlin Richard E. , Shpall Elizabeth J. , Rezvani Katayoun
Published: December 2023

Despite the high response rates of chimeric antigen receptor (CAR) - T cells targeting the B-cell maturation antigen (BCMA) for multiple myeloma (MM), patients eventually still relapse. At this time there is no clear subsequent therapy. Furthermore, CAR T-cell therapy is associated with significant toxicities such as cytokine release syndrome and immune effector cell-associated neurotoxicity syndrome.

CD70 is a normally transiently expressed on activated T-cells, B-cells, or dendritic cells, but have been found to be upregulated on many hematologic and solid cancer cells such as acute myeloid leukemia, triple negative breast cancer, and renal cell carcinoma. However, CD70 as a potential target for treatment of multiple myeloma, including patients who have progressed on BCMA targeted therapies, has not been fully investigated.

We assessed bone marrow aspirates from MM patients by flow cytometry and demonstrated that CD70 was expressed on the majority of myeloma cells, and on average, relapsed myeloma patients had higher expression of CD70. In addition, using immunohistological staining, we found that CD70 was expressed on 10 of 10 patients who had progressed on BCMA targeted therapies (seven who had progressed on belantamab mafodotin, an antibody drug conjugate, three who had progressed on idecabtagene vicleucel, a BCMA CAR-T), suggesting that CD70 could be a viable target in patients who have failed BCMA targeted therapy.

As natural killer (NK) cells are a promising off-the-shelf treatment with minimal toxicity as compared to CAR-T cells, we generated CD70-targeting CAR NK cells for the treatment of MM. We found in the chromium cytotoxicity assay that CD70 CAR NK cells had higher killing as compared to non-transduced (NT) NK cells at multiple effector-to-target ratios against MM1s, a myeloma cell line confirmed to express CD70. There was no difference in killing when co-cultured with H929, a myeloma cell line lacking CD70 expression, demonstrating the specificity of the CD70 CAR NK cell. The enhanced killing was also confirmed in an incucyte assay in which the CD70 CAR NK cells cleared tumor faster than NT NK, even in MM1s in which BCMA was knocked out using CRISPR (simulating BCMA antigen loss). When tested in a myeloma mouse model, the CD70 CAR NK exhibited greater tumor control as shown in Figure 1, when compared to NT NK, which lead to increased survival of the mice. Similar results were seen in a mouse model using BCMA KO myeloma cells.

In summary, we were able to demonstrate that CD70 can be a feasible target for the treatment of multiple myeloma, including in patients who have failed BCMA targeted therapy. Based on these results, we have initiated a Phase I/II clinical trial (NCT05092451) that is currently recruiting.
Successful Treatment Outcomes of Hematopoietic Stem Cell Transplantation with Reduced-Toxicity Conditioning Regimen Incorporating Treosulfan in Pediatric Patients with XIAP Deficiency
Authors: Suh Jin Kyung , Im Ho Joon , Kang Sung Han , Kim Hyery , Choi Eun Seok , Koh Kyung-Nam
Published: December 2023

Background

X-linked inhibitor of apoptosis (XIAP) defciency is an inherited primary immunodefciency characterized by chronic infammasome overactivity and associated with hemophagocytic lymphohistiocytosis (HLH) and infammatory bowel disease (IBD). Although hematopoietic stem celltransplantation (HSCT) is the only curative therapy, the outcomes of HSCT for XIAP deficiency remain unsatisfactory.

Methods

We have performed HSCT with treosulfan based RTC regimen for pediatric patients with non-malignant disorders since January 2016, and reviewed the medical records of patients with XIAP deficiency who underwent treosulfan based HSCT to investigate outcomes of HSCT for patients with XIAP deficiency.

Results

Since January 2016, 4 patients with XIAP deficiency had undergone HSCT using RTC regimen consisted of fludarabine (150-180 mg/m 2), treosulfan (42 g/m 2), rabbit ATG (5-7.5 mg/kg), and thiotepa (10 mg/kg). All patients were male and received HSCT (2 from URD and 2 from HFD) at median 13.7 years old (range, 12.8-15.3), median 2 years (range, 0.5-3.9) after diagnosis of XIAP. They all achieved engraftment and complete donor chimerism at the time of last evaluation. One patient who received HFD HSCT developed acute GI GVHD grade 3 and VOD within 1 month from HSCT, which was recovered after treatment with systemic steroid and defibrotide. Except the patient, there was no fatal acute regimen related toxicity ≥ CTCAE grade 3. Two patients developed chronic GVHD: one had limited disease which was resolved after systemic steroid therapy and the other had extensive disease. During the median 43 months (range, 33.1-63.0) follow-up, all survived without disease recurrence.

Conclusions

HSCT with treosulfan based RTC regimen is promising treatment option for pediatric patients with XIAP deficiency, with successful engraftment and low treatment-related toxicity. However, further studies including a larger multicenter trial and measures to refine the regimen are mandate to verify efficacy and safety of this regimen.
Renal Failure in Patients with Newly Diagnosed Multiple Myeloma Eligible for Autologous Stem Cell Transplant in LATAM - on Behalf of Gelamm
Authors: Martinez-Cordero Humberto , Fuentes Lacouture Maria Cynthia , Von Glasenapp Seisha Alana , Peña Camila , Riva Eloisa
Published: December 2023

Background

Renal failure (RF) is a well-known complication in patients with multiple myeloma (MM). It is reported that RF occurs in 20% to 50% of patients at the time of diagnosis. Approximately 10% will require hemodialysis. It is important to consider that RF determines a worse short-term prognosis and decreased survival in these patients, especially for those requiring dialysis.

Methods

An international, multicenter, retrospective observational cohort study was conducted. Data were collected from the GELAMM retrospective Registry. We analyzed demographical and clinical characteristics, and overall survival (OS) of newly diagnosed MM patients without RF, patients with RF not requiring hemodialysis (HD), and patients with RF requiring HD. The logistic regression model was used to identify risk factors for worse outcomes.

Results

We included 1104 NDMM patients ≤ 65 years old from GELAMM registry: 854 patients without renal failure, 161 patients (15%) with RF not requiring HD, and 89 patients (8%) requiring HD. There were no differences in terms of demographic and clinical characteristics. With a median follow-up time of 29 months, the median OS was not reached (NR) in patients without RF and in patients with RF without HD requirement, however, it was 43 months (27 - 58 months) in patients with renal failure requiring HD (p=<0.001). The HR for mortality in patients with RF without HD was 1.75 (1.27 -2.41) and for RF and HD 3.49 (2.47 - 4.93) (p< 0.001). The one-year mortality among the three groups was 7% for patients without RF, 12% in patients with RF not requiring dialysis, and 30% in patients with RF requiring dialysis. The multivariate analysis showed that having proceeded with ASCT as a favorable prognostic factor HR 0.238 (0.129-0.521) p< 0.001 and high-risk cytogenetics as an unfavorable prognostic factor HR 2.936 (1.242-6.922) p=0.014.

Discussion

Renal failure confers a worse prognosis in Latin American patients with newly diagnosed multiple myeloma, especially for those who require dialysis, and increases one-year mortality more than 4 times. The use of ASCT decreases the risk of worse time-dependent outcomes, and high-risk cytogenetics increases the risk of death.
Avatrombopag Is Effective in First-Line Treatment of Severe Aplastic Anemia: Early Outcomes of a Prospective Study
Authors: Zhao Xin , Li Jianping , Fan Huihui , Jing Liping , Zhang Li , Zhou Kang , Yang Wenrui , Li Yuan , Yang Yang , Ye Lei , Peng Guangxin , Xiong Youzhen , Shi Jun , Zhang Fengkui
Published: December 2023

Acquired severe aplastic anemia (SAA) is a life-threatening bone marrow failure disease and its pathogenesis involves T cell-mediated autoimmunity. Antithymocyte globulin (ATG) and cyclosporine based immunosuppressive therapy (IST) is effective for patients not eligible for hematopoietic stem cell transplant, and the response rate is 60-70%. Complete hematological response (CR) predicts better survival of patients received IST treatment, but only a few patients achieve CR within 6 months (5% at 3 months and 12-17% at 6 months). Thrombopoietin receptor agonist (TPO-RA) eltrombopag improves the overall response (OR) rate and CR rate of SAA patients when combined with IST, but its hepatotoxicity is still worrying.

Avatrombopag is another small-molecule TPO-RA without hepatic toxicity. We previously found that SAA patients received avatrombopag and IST as first-line treatment showed both higher CR rate and OR rate at 3 and 6 months. To validate our findings, a prospective, single-arm study of avatrombopag and IST for newly diagnosed SAA was carried out (NCT05720234).

There were 25 patients enrolled in the study by June 31, 2023, including 21 SAA patients and 4 very severe aplastic anemia patients. Patients were diagnosed as AA within 6 months and received ATG, cyclosporine and avatrombopag as first-line treatment. Avatrombopag was administered at the first day of ATG initiation at dose of 60mg per day for patients weight ≥ 50kg and 40mg per day for patients weight < 50kg. ATG was given for 5 consecutive days. Cyclosporine was given orally at dose of 3.5-5mg/kg.d.

There were 15 males and 10 males, at a median age of 37 years old. Eighteen patients achieved the end-point of 3 months and 10 patients achieved end-point of 6 months. There were 15 patients achieved hematologic response at 3 months (83.3%), including 5 complete responders (27.8%) and 10 partial responders. The overall response rate at 6 months was 80%, and CR rate was 60%. One patient evolved to hemolytic paroxysmal nocturnal hemoglobinuria at 3 months, with normal count of platelet and white blood cell.

In conclusion, the addition of avatrombopag to IST as first-line treatment showed earlier hematologic response and higher complete response rate for patients with SAA.