bluuebunny/crossref_metadata_2025 huggingface dataset:
Insights on Sickle Cell Disease Therapies in Sierra Leone
Authors: Willard Rachele E. , Buscetta Ashley , Ramirez Hasmin C. , Jaja Cheedy , Bonham Vence L.
Published: December 2023

Introduction: Ongoing sickle cell disease (SCD) gene therapy clinical trials in the United States and other high resource countries have shown promising results, and several therapies are under U.S. Food and Drug Administration review in 2023. Despite these advances, the current evidence-based practices are not accessible to much of the world's sickle cell population. As treatment advances are made for SCD, the integration of these therapies in sub-Saharan African countries-which have the highest prevalence of SCD-requires a global partnership. Sierra Leone ranks 9 th in burden in sub-Saharan Africa with annual SCD births between 4000-5000. Efforts are underway in Sierra Leone to improve awareness and healthcare infrastructure for the SCD population; however, the country still lacks a national screening program for diagnosis and faces challenges in providing comprehensive care across the lifespan. This study examines adult SCD patients' knowledge of current evidence-based care and novel gene therapies in Sierra Leone.

Methods: In 2021-2022, a cohort (n=123) of adults living with SCD in Sierra Leone were surveyed as part of the INSIGHTS Study (NCT02156102). A mixed-methods recruitment strategy was used, including via public service announcements hosted on local radio stations, information disseminated via a WhatsApp group, and snowball sampling. Recruitment was conducted in two settings, one urban (Freetown) and one rural (Kono). Participants completed this cross-sectional study from September 2021-February 2022. Questions were asked regarding participants' perspectives and knowledge of hydroxyurea, the current standard of care treatment for SCD, and novel experimental gene therapy.

Results: The mean age of respondents was 23.6 years (range 18-63). The majority of respondents were female (65.6%), educated at or above the senior secondary school level (78.7%), and students (70.3%). Respondents predominantly had the HbSS genotype (89.4%). Most study participants lacked awareness of hydroxyurea and gene therapy (93.5% had never heard of hydroxyurea; 95.1% had never taken hydroxyurea; 94.8% had never heard of gene therapy for SCD treatment) (Table 1). Despite this gap in knowledge, the majority (95.0%) agreed that genetic research is a necessary way to learn about treatments for SCD (Table 1).

Discussion: Our results highlight disparities in current standard of care treatment and knowledge in a country with a high prevalence of SCD. While our cohort indicated a favorable attitude toward SCD research and a willingness to participate, these results highlight a significant barrier for successful implementation of current treatment and the adoption of curative genetic therapies in limited resource settings like Sierra Leone. Given the significant lack of knowledge around hydroxyurea, it is important that public health initiatives and research should be directed toward both established standard of care and creating awareness of experimental therapies.
Lipid Nanoparticle Library Towards Development of Next Generation Genomic Medicines
Authors: Jain Nikita , Nazaripour Sedigheh , Chen Zhengyu , Abraham Suraj , Yee Leanna , Sadat Sams , Mehar Namala Gayatri , Son Helena “” , Chemmannur Sijo , Tickner Kobe , Anantha Malathi , Sharma Ruchi , Abbina Sri , Thambatti Seetalakshmi , Mayya Vinay , Soon Emily , Paquette Jay , Thomas Anitha , Nguyen Viet
Published: December 2023

Ionizable amino lipids are a major constituent of the lipid nanoparticles for delivering nucleic acid therapeutics (e.g., DLin-MC3-DMA in ONPATTRO®, ALC-0315 in Comirnaty®, SM-102 in Spikevax®). Scarcity of lipids that are suitable for cell therapy, vaccination, and gene therapies continue to be a problem in advancing many potential diagnostic/therapeutic/vaccine candidates to the clinic.

Herein, we describe the development of novel ionizable lipids to be used as functional excipients for designing vehicles for nucleic acid therapeutics/vaccines in vivo or ex vivo use in cell therapy applications. We first studied the transfection efficiency (TE) of LNP-based mRNA formulations of these ionizable lipid candidates in primary human T cells and established a workflow for engineering of primary immune T cells. We then adapted this workflow towards bioengineering of CAR constructs to T cells towards non-viral CAR T therapy. Lipids were also tested in rodents for vaccine applications using self-amplifying RNA (saRNA) encoding various antigens. We have then evaluated various ionizable lipid candidates and their biodistribution along with the mRNA/DNA translation exploration using various LNP compositions. Further, using ionizable lipids from the library, we have shown gene editing of various targets in rodents.

We believe that these studies will pave the path to the advancement in nucleic acid based therapeutics and vaccines, or cell & gene therapy agents for early diagnosis and detection of cancer, and for targeted genomic medicines towards cancer treatment and diagnosis.
The Impact of Immune Function on Hematopoietic Reconstitution and Infection in Patients with Lymphoma after Autologous Hematopoietic Stem Cell Transplantation
Authors: Hu Renzhi , Liu Jun , Liang Xiping , Zhang Wenjun , Zhou Yixing , Li Qiying , Liu Yao
Published: December 2023

Objective

To investigate the impact of immune function on hematopoietic reconstitution and infection in lymphoma patients after autologous hematopoietic stem cell transplantation (auto-HSCT).

Methods:

A total of 20 patients with lymphoma who underwent auto-HSCT in Chongqing University Cancer Hospital from January 2021 to December 2021 were retrospectively analyzed. According to whether the number of CD4+T cells was greater than or equal to 200 cells /ul, the patients were divided into normal immune function group and immunocompromised group. The hematopoietic reconstitution of the two groups was observed, including the time of neutrophil engraftment and platelet engraftment, as well as the occurrence of infection, including the pathogen could not be identified in some patients with neutropenia, it was necessary to judge according to physical examination, laboratory and imaging examination or empirical antimicrobial treatment was effective.

Results

1. Immunocompetent and immunocompromised patients included 10 patients, respectively. The durations of neutrophil engraftment were 9.5 (7-12) days and 12 (8-15) days (P = 0.007), and the durations of grade 3 neutropenia were 1.5 (0-4) days and 3 (1-7) days (P = 0.030), respectively. The duration of grade 4 neutropenia was 9 (5-16) days and 11 (6-20) days, respectively (P = 0.010). There was no significant difference in platelet engraftment time and incidence of grade IV anemia between the two groups (P >0.05). 2. The incidence of febrile neutropenia in immunocompetent group and immunocompromised group was 60% and 80%, respectively, and the median duration of febrile neutropenia was 3.5 (1-10) days and 5 (1-20) days, respectively (P = 0.005). 3. The incidence of infection in immunocompetent group and immunocompromised group was 20% and 30%, and the incidence of bloodstream infection was 20% and 30%. 4. The length of hospital stay in immunocompetent group and immunocompromised group was 31.5 (23-43) days and 37 (25-75) days, respectively (P<0.001).

Conclusion

The immune function of lymphoma patients before transplantation has a certain impact on hematopoietic reconstitution and infection. Normal immune function is superior to immunocompromised in terms of neutrophil engraftment, shortening the duration of febrile neutropenia, and the incidence of infection after transplantation.
KSHV/HHV-8 Associated Multicentric Castleman Disease in 80 HIV Negative Patients
Authors: Stemmler Romain , Malphettes Marion , Galicier Lionel , Gerard Laurence , Meignin Veronique , Boutboul David , Oksenhendler Eric
Published: December 2023

Introduction : KSHV/HHV-8 associated multicentric Castleman disease (HHV-8 MCD) is a rare lymphoproliferative disorder associating lymphadenopathy, splenomegaly, fever and inflammatory symptoms with high serum CRP, hypergammaglobulinemia and high plasma HHV-8 DNA viral load. HHV-8 MCD mainly affects HIV infected patients but can also be observed in HIV negative patients born in countries with high HHV-8 prevalence or in men who have sex with men. In a large cohort of HIV negative patients, we sought to describe patient and disease characteristics and to compare them with those observed in a large cohort of HIV-infected patients with HHV8 MCD.

Methods : We analysed a cohort of 80 HIV negative patients with HHV8+ MCD and compared them to 228 HIV-infected patients with HHV8+ MCD. These patients have been prospectively enrolled in the national registry for Castleman disease held by the Reference Center for Castleman Disease in Paris, over a 30-year period.

Results : Most of these patients (67,5 %) originated from Sub saharan or Northern Africa. The baseline characteristics were similar in HIV- and HIV+ patients although HIV negative patients were older at MCD diagnosis (median, 67 vs 43 years) with 20% of the patients above 75 years, and the male predominance was slightly lower in the HIV negative population (74% vs 82%). They also presented less frequently with previous Kaposi sarcoma lesions (34% vs 52%). Biological markers including cytopenia, serum CRP and gamma globulins levels as well as blood HHV8 DNA viral load were very similar in both groups, although MCD attacks in HIV negative patients were rather less agressive than in HIV-infected patients. Of notice, the median CD4 + T cell count was normal in the HIV negative group (600 x10 6/L)

The incidence of lymphoma during follow-up remains high but very similar in both groups : 3,12 /100 pt.y. in HIV negative patients and 3,10 /100 pt.y. in HIV-infected patients

The median overall survival (OS) was 8,8 years (95%CI, 5,5-15) in HIV- patients and 20,6 years (95%CI, 6,3-24,1) in HIV-infected patients (p=0.19). Although the follow-up was shorter in HIV negative patients (24 vs 56 months), the 5-year probability of survival (79,2%; 95%CI 61,7-89,4) was very similar to that observed in HIV-infected patients (81,2% ; CI95% 74,6-86,2). Death was related to HHV8 complications or treatment in 6 /15 (40%) HIV negative patients and in 31 /57 (54%) HIV-infected patients. The slightly higher proportion of deaths not related to HHV8 in the HIV negative population might be explained by the older age of these patients.

Conclusions : Using a large cohort of HIV negative patients, we could show that HHV8 MCD in this population is very similar to that observed in HIV-infected patients. Despite the fact that these patients were older at diagnosis, the overall survival was similar to that observed in HIV-infected patients, at least at 5-year from diagnosis.
Comparison of Two Venetoclax-Based Regimens As Salvage Therapies Bridged to Allogeneic Hematopoietic Cell Transplantation in Patients with Relapsed/Refractory Acute Myeloid Leukemia
Authors: Ye Yiling , Chen Yanqiu , Yu Guopan , Weng Guangyang , Liu Qifa , Jin Hua
Published: December 2023

Introduction ：

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective treatment for relapsed or refractory acute myeloid leukemia (R/R-AML) patients. However, it is not clear about appropriate salvage therapy bridged to allo-HSCT for R/R-AML patients. In this study, we compared the outcomes of R/R-AML patients treated with venetoclax plus hypomethylating agents (VEN-HMA) regimen or venetoclax plus azacitidine and homoharringtonine (VAH) regimen as salvage therapies bridged to allo-HSCT.

Methods ：

A total of 105 R/R-AML patients treated with VEN-HMA regimen or VAH regimen bridged to allo-HSCT between September 2018 and February 2023 were enrolled. The outcomes including survival, relapse and NRM after allo-HSCT were analyzed in two groups.

Results ：

Forty-four patients were treated with VEN-HMA regimen, and 61 with VAH regimen. Seventy-four patients had primary refractory AML and 31 relapsed AML, including 19 relapsed after chemotherapy and 12 relapsed after allo-HSCT. Thirty-one (70.5%) patients achieved composite complete remission (CRc) in VEN-HMA group including 20 (45.5%) with minimal residual disease (MRD) negative, as well as 52 (85.2%) with CRc including 40 (65.6%) with MRD negative in VAH group (P=0.09). Twenty-five (56.8%) patients were stratified with adverse risk in VEN-HMA group and 45 (73.8%) in VAH group according to the 2022 European Leukemia Net (ELN) classification (P=0.07). The 2-year overall survival (OS) and event-free survival (EFS) in VAH group were higher (78.4% vs 58.7%, P=0.03; 68.3% vs 38.7%, P=0.003), as well as 2-year cumulative incidence of relapse (CIR) was lower than VEN-HMA group (23.2% vs 42.4%, P=0.04). Among patients with CRc, the 2-year OS, EFS and CIR was 67.5%, 48.9% and 37.0% in VEN-HMA group, as well as 82.5%, 72.5% and 23.6% in VAH group (P=0.15, 0.05 and 0.24). Among patients with MRD negative, the 2-year OS, EFS and CIR was 75.5%, 55.5% and 31.6% in VEN-HMA group, as well as 90.8%, 80.0% and 17.5% in VAH group (P=0.15, 0.10 and 0.28). Among patients with adverse risk, the 2-year OS and EFS in VAH group were higher (79.3% vs 47.7%, P=0.01; 70.9% vs 24.1%, P=0.0002), as well as 2-year CIR was lower than VEN-HMA group (17.4% vs 45.8%, P=0.01). Non-relapse mortality (NRM) was similar between VEN-HMA group and VAH group (18.9% vs 8.5%, P=0.12).

Conclusion ：

VAH regimen is an effective salvage therapy bridged to allo-HSCT for R/R-AML patients compared with VEN-HMA regimen.

Keywords ：

Venetoclax, Azacitidine, Homoharringtonine, Relapsed/Refractory AML, Allogeneic Hematopoietic Cell Transplantation.
Three-Day Decitabine Combined with Venetoclax in Patients with Newly Diagnosed or Relapsed/Refractory AML
Authors: Suo Xiaohui , Bai Guanchen , Zhao Xingli , Zhang Congcong , Li Yinling , Gao Sifeng , Zhang Jilei , Lu Xinxiao , Yuan Linyu , Mi Yingchang , Liu Kaiqi
Published: December 2023

INTRODUCTION

Venetolax (VEN) combined with azacytidine (AZA), decitabine (DEC), and low-dose cytarabine has been the first-line therapy for newly diagnosed (ND) AML patients who are elderly or unfit for intensive chemotherapy. Previous reports shown that ten-day DEC with VEN (DEC 10-VEN) was an effective therapy for newly diagnosed elderly patients. Though higher remission rate in ND AML compared five-day DEC with VEN, DEC 10-VEN induced prolonged myelosuppression. Therefore, optimizing the combination of DEC with VEN is still being explored.

In this study, we retrospectively collected and analyzed the clinical data of three-day DEC combined with VEN (DEC 3-VEN) ( Tianjin-Handan regimen) in twelve patients with ND or relapsed/refractory (R/R) AML who were elder or ineligible for intensive chemotherapy to evaluate the efficacy and safety of the regimen.

METHODS

Retrospectively collected and analyzed the clinical data of twelve ND or R/R AML patients treated with three-day DEC combined with VEN as induction therapy between July 2022 and June 2023 at 4 hospitals in china.

The induction regimen consisted of oral Ven (100mg Day 1; 200mg Day 2; 400mg Day 3-9 or 3-14), intravenous DEC (20mg/m²/q8h, days 4-6), sorafenib were administered (600mg days 8-14) in relapsed/refractory patients with FLT3/ITD. The primary objective was composite complete response rate (CR + CRi) and the secondary objectives was safety.

RESULTS

Until June 2023, twelve patients with de novo or relapsed/refractory AML were treated with three-day DEC combined with VEN as induction therapy. The median age of the twelve patinets was 67 years (range, 32-76), with 58.3% (7/12) male. Seven (58.3%) patients were ND AML, and five patients (41.7%) were R/R AML. The clinical data and treatment information of the patients are shown in table 1.

The composite complete response rate (CR + CRi) after one cycle of induction was 100% (CR 10/12, CRi 2/12). For patients who achieved CR or CRi, MRD negative rate was 77.8% (7/9) by flow cytometry. None of patients died during the induction therapy.The median time to recovery of the absolute white blood count (WBC) to ≥1.0 × 10 9/L and the platelet count to ≥20 × 10 9/L after induction was 13 (range: 7-25) and 15 days (range: 0-27), respectively.

CONCLUSION:

DEC 3-VEN is a highly effective and safe induction therapy for ND and R/R AML patients who are elderly or unfit for intensive chemotherapy. Further study with more patients will be updated.

【Keywords】Venetoclax; decitabine; AML; induction treatment.
Evaluation of Innovative Laboratory Tests to Predict a Thrombotic Phenotype in a Family with Dysfibrinogenemia and a Novel FGG Mutation
Authors: Monard Amaury , Moenen Floor , Castoldi Elisabetta , De Simone Ilaria , Van Der Meijden Paola , Wichapong Kanin , van Doorn William , Spada Stefano , Hellenbrand Dave , Stork Alexander , Ten Cate Hugo , Beckers Erik AM , Henskens Yvonne
Published: December 2023

Background:

Hypodysfibrinogenemia is a rare hereditary fibrinogen disorder characterized by quantitative and qualitative fibrinogen defects. These fibrinogen defects can cause thrombotic and hemorrhagic phenotypes. Unfortunately, predicting the phenotype in a specific patient is often not possible with routine coagulation tests.

Aims:

To characterize the phenotype and the genetic profile of a family with hypodysfibrinogenemia and to investigate the ability of innovative tests to predict bleeding and/or thrombotic phenotypes in asymptomatic family members.

Methods:

The proband, a 60-year-old woman with both bleeding and thrombotic complications who is currently on DOAC treatment, and two daughters were referred to our Hemophilia Treatment Center (HTC) for phenotypical and genotypical analysis of a congenital fibrinogen disorder (CFD). Extensive laboratory testing was done, as well as DNA-sequence analysis and molecular modelling. Thrombin generation and microfluidic testing were also performed to investigate their ability in phenotype prediction.

Results:

Fibrinogen activity and antigen levels led to the diagnosis of dysfibrinogenemia in the proband and hypodysfibrinogenemia in both daughters. In all three cases, the same heterozygous missense mutation in the FGG gene was uncovered. This likely pathogenic mutation leads to the p.(Tyr375Cys) amino acid change. Molecular modeling predicted possible conformational changes or covalent dimerization of the fibrinogen molecule. Thrombin generation was elevated in one daughter. Microfluidic testing showed enhanced fibrin formation in both daughters, regardless of the coagulation trigger.

Conclusion:

We described a family with hypodysfibrinogenemia in whom a novel heterozygous missense mutation in the FGG gene was found, possibly leading to conformational changes or covalent dimerization of the fibrinogen molecule. Furthermore we showed that microfluidic testing and thrombin generation can indicate a thrombotic phenotype in these patients, not detected with routine coagulation tests.
Elevated Functional Hematopoietic Stem and Progenitor Cells in the Peripheral Blood of G-CSF-Mobilized Dnmt3a-Mutant Mice
Authors: Young Kira A. , Nye Griffin J. , Trowbridge Jennifer J.
Published: December 2023

Clonal hematopoiesis (CH) arising from somatic mutations in hematopoietic stem cells (HSCs) is associated with increased risk of developing several non-hematologic and hematologic diseases. In the context of stem cell transplantation, donor HSCs carrying CH-associated somatic mutations can be passed from donor to recipient during stem cell transplantation. Most literature supports that there is no negative impact on recipient survival or outcomes after allogeneic stem cell transplantation (allo-HSCT) from donors with CH. However, it is less clear how allo-HSCT donors with CH respond to common mobilization methods such as Neupogen (G-CSF). As hematopoietic stem and progenitor cells (HSPCs) with CH mutations have been found to respond to external stimuli such as inflammation in a manner distinct from wild-type HSPCs, it is plausible that CH-mutant HSPCs also distinctly respond to G-CSF mobilization. It is important to understand whether the process of mobilization differentially enriches for CH-mutant HSPCs in the mobilized peripheral blood product. To address this gap in knowledge, we used a murine model of Dnmt3a-mutant CH. After five consecutive days of Neupogen (G-CSF) administration, modeling the procedure for human HSC mobilization, we observed that Dnmt3a-mutant mice had significantly increased white blood cell, neutrophil and monocyte counts compared to control mice. We observed a significant increase in cell surface marker-defined HSPCs in the peripheral blood and spleen of G-CSF-treated Dnmt3a-mutant mice compared to G-CSF-treated control mice. Using in vitro colony-forming unit assays, the peripheral blood of G-CSF-treated Dnmt3a-mutant mice was found to contain a greater number of functional hematopoietic progenitors compared to G-CSF-treated control mice. To determine if the peripheral blood of G-CSF-treated Dnmt3a-mutant mice had a greater abundance of functional HSCs, we transplanted equal volumes of mobilized blood from Dnmt3a-mutant and control mice into primary and secondary lethally irradiated wild-type recipient mice. We observed increased long-term engraftment of mobilized blood from Dnmt3a-mutant mice compared to control mice in both primary and secondary recipient animals. At a mechanistic level, HSC mobilization is facilitated by myeloid cells that respond directly to G-CSF via the G-CSF receptor ( Csf1r). In Dnmt3a-mutant mice, we observed increased Csf1r expression in myeloid progenitor and mature myeloid cells compared to control mice, providing a putative mechanism by which increased mobilization is enabled. Together, our data support differential mobilization potential of Dnmt3a-mutant compared to control HSCs in response to G-CSF, which may have implications in the context of allo-HSCT.
Clinical Analysis of Metagenomic Sequencing of Plasma Microbial Cell-Free DNA for Infected Patients in SAA
Authors: Jing Liping , Fan Huihui , Yang Wenrui , Xiong Youzhen , Zhang Li , Zhao Xin , Zhang Fengkui
Published: December 2023

Backgroun d: As a high-risk group of clinical infection, patients with severe aplastic anemia (SAA) have high mortality due to agranulocytosis or immune function impairment. The current conventional detection methods such as blood culturetissue chulture etc. often cannot identify pathogenic bacteria. We evaluate the diagnostic performance of metagenomic next-generation sequencing (mNGS), and explore the application value of mNGS in infected patients in SAA, in order to provide help for precision therapy.

Methods: From Ocb 2020 to Dec 2022 in our center, we enrolled 70 SAA patients with infection and investigated the usefulness of mNGS of plamsa or tissue DNA for identification of infectious pathogens. The consistency of the results of mNGS and the results of routine etiological detection (blood culture(BC), swab/tissue culture) with clinical infection diagnosis and the influence of the results of mNGS on clinical infection treatment were compared.

Results: 70 SAA patients of mNGS were detected, including plasma-mNGS 66 cases and tissue-mNGS 4 cases. Among all patients, 48 cases(68.6%) were positive. In comparison to blood culture, the positive and negative agreement of mNGS were 100% (13/13) and 38.6% (22/57), respectively. In 48 mNGS positive cases, 36 patients (75%) underwent antimicrobials adjustment, resulting in positive impact on 30 patients. Fungi were detected in 29 cases, the detection rate was 41.4% (29/70) by mNGS, otherwise 2.8% (2/70) by BC. Of the 70 patients with SAA, 55 infections eventually impproved (78.6%).

Conclusion: The positive rate of mNGS detection and the consistent rate with clinical diagnosis are higher than that of conventional etiological detection, which provides a faster and more sensitive detection method for infected patients in SAA.

【 Key words 】 Metagenomic Sequencing ; Plasma Microbial Cell-free DNA; Aplastic anemia; Infection; pathogen diagnosis
Familial Thrombocytopenia-Associated Germline ETV6 P214L Mutation Results in XPO1-Mediated Nuclear Export
Authors: McCONVILLE Michael , Thomas Toby , Valadez Catherine , Beckner Ryan , Chook Yuh Min , Chung Stephen , Liszczak Glen
Published: December 2023

In recent years, a number of heterozygous germline mutations in ETV6 have been associated with familial thrombocytopenia and predisposition to hematologic malignancies. While most of these missense mutations occur in the DNA-binding-domain of ETV6, the recurrent P214L mutation is unique in that it occurs in the central disordered region of the protein. The ETV6 P214L mutation has previously been shown to disrupt the nuclear localization of ETV6 and the ability of ETV6 to act as a transcriptional repressor. Unexpectedly, we have found that the P214L missense mutation creates a recognition motif for the nuclear export protein XPO1. We have demonstrated in cellular and protein reconstitution assays that XPO1 directly engages the P214L but not the wild-type ETV6 construct. Consistent with this mechanism, we are able to fully rescue the nuclear localization of ETV6 P214L in mammalian cells with small molecule XPO1 inhibitors. We have also found that once localized to the nucleus, the P214L mutant construct exhibits functional consequences indistinguishable from wild-type ETV6 in Ba/F3 transformation assays.

When looking at the evolutionary conservation of the ETV6 amino acid sequence involved in the XPO1 recognition motif, we observed that the synonymous P216L mutation in the mouse ETV6 ortholog does not create a nuclear export signal. In order to study XPO1-mediated export of ETV6 P214L and its impact on hematopoiesis, we have generated a mouse model in which two point mutations (P216L and S211I) have been introduced into a single endogenous ETV6 allele, which precisely matches the sequence of this region in human P214L carriers. Using CRISPR-Cas9, we generated 3 transgenic mouse lines: Etv6 P216L, Etv6 S211I, and Etv6 S211IP216L. Immunofluorescence of fixed bone marrow shows that ETV6 is only mislocalized in megakaryocytes from ETV6 S211I P216L mice. Complete blood counts revealed a significant decrease in platelet counts in ETV6 S211I P216L heterozygotes (485 x 10 3/µL) compared to ETV6 S211I heterozygotes (1140 x 10 3/µL, p<0.0001), ETV6 P216L heterozygotes (1065 x 10 3/µL, p<0.0006), and wild-type littermates (1254 x 10 3/µL, p<0.0001). Flow cytometry phenotyping of bone marrow from the three transgenic lines and wild-type littermates shows a significant decrease in the absolute numbers of hematopoietic stem cells (HSCs) and megakaryocyte precursors (MkPs) in ETV6 S211I P216L mice.

In summary, we have determined that XPO1-mediated nuclear export is the mechanistic basis for ETV6 P214L mislocalization. We developed a mouse model that recapitulates this export signal in the endogenous ETV6 allele and found that this model demonstrates a thrombocytopenic phenotype as seen in human patients. Reductions in HSCs and MkPs suggest both a decreased fitness of HSCs, as well as dysregulated megakaryopoiesis in this model.
Underrepresentation of Racial and Ethnic Minorities in Registrational Chimeric Antigen Receptor (CAR) T-Cell Therapy Trials in Hematologic Malignancies: An FDA Analysis
Authors: Mahat Upendra , Seam Pamela , Bouchkouj Najat , Sharma Poornima , Peredo-Pinto Helka , Elmacken Mona , Kaushal Megha , Wang Shu , Xu Zhiheng , Jia Yuxia , Richardson Nicholas , Kanapuru Bindu , Kasamon Yvette , Gormley Nicole
Published: December 2023

Introduction: Equitable representation of diverse racial and ethnic groups in oncology clinical trials continues to be a major challenge. Numerous initiatives designed to enhance diversity in oncology clinical trials are ongoing, including efforts at the U.S. Food and Drug Administration (FDA) such as the Oncology Center of Excellence's Project Equity. Given the multiple recent FDA approvals of CAR T-cell therapies for relapsed or refractory hematologic malignancies, we evaluated the enrollment rates of racial and ethnic groups in the CAR T-cell registrational trials submitted to the FDA.

Methods: We conducted a pooled, patient-level analysis of all registrational trials of CAR T-cell therapies submitted to the FDA to support approval for hematologic malignancies between 2017 and 2021. Datasets from approved licensing applications were pooled, and demographics and efficacy outcomes were analyzed descriptively.

Results: A total of 13 registrational trials involving 6 autologous CAR T-cell products were analyzed, comprising 1,319 patients with hematologic malignancies. The median age at enrollment was 60 years (range: 3 to 86 years). Of the 1,319 patients, 81% (1068) were White, 5% (66) were Black, 5% (65) were Asian, 4% (48) were reported as Other, and 8% (110) were Hispanic or Latino. Race and ethnicity information was not reported for 5% (72) and 10% (127) of patients, respectively. The majority of the patients enrolled in the registrational trials were from the United States (80%), followed by Europe (11%) and Australia (2%). The hematologic malignancies included large B cell lymphoma (59%), multiple myeloma (15%), follicular lymphoma (13%), B cell acute lymphoblastic leukemia (9%), and mantle cell lymphoma (5%). Response rates by race, ethnicity, and histology are shown in Table 1 and Table 2.

Conclusions: Despite the majority of patients being enrolled in the U.S., racial/ethnic subgroups other than White and non-Hispanic/Latino were underrepresented in CAR T-cell registrational trials. This highlights the need to understand and overcome the continued barriers to CAR T-cell clinical trial enrollment across racial and ethnic groups. Although clinically meaningful efficacy was observed across these racial/ethnic groups and diseases, more data are needed to evaluate if efficacy outcomes vary across these subgroups. Work to describe safety and tolerability, particularly cytokine release syndrome (CRS) and neurotoxicity, in these racial and ethnic subgroups is ongoing. More effort is needed to better enhance diversity in CAR T-cell trials, especially given the expanding use of CAR T-cell therapies in the treatment of patients with hematologic malignancies.
Tern-701 (HS-10382) Is a Potent Inhibitor of BCR::ABL1 and Is Synergistic with Active Site Tyrosine Kinase Inhibitors
Authors: M Parsons Benjamin , Jasper Jeffrey R. , Jones Christopher
Published: December 2023

CONCLUSIONS

Although tyrosine kinase inhibitors (TKIs) targeting the ATP-binding site of the BCR::ABL1 oncoprotein are effective therapeutics for chronic myeloid leukemia (CML), patients often develop drug resistance due to ATP-site mutations that inhibit drug binding. TERN-701 is a potent and highly selective inhibitor of BCR::ABL1 that is designed to bind the allosteric myristoyl site on the kinase, circumventing resistance to active site mutations, with potential for synergistic combination with active site TKIs.

Competition binding screens were used to assess the selectivity of TERN-701 on more than 450 targets spanning the human kinome. The potency of TERN-701 against the proliferation of wild type and mutant CML cell lines, as well as a panel of 102 cancer cell lines, was assessed using CellTiter-Glo®. Synergy between TERN-701 and active site TKIs was assessed in vitro with interactions quantified using Bliss, combination, and curve-shift analyses. Prospective mutagenesis experiments were conducted by treating BaF/3 cells engineered to overexpress the BCR::ABL1 kinase with various concentrations of both TERN-701 and the TKI imatinib, followed by observation of clonal outgrowth of treated cells over time.

We have previously shown that TERN-701 is a potent inhibitor of native CML cell lines such as K562. Here, we show that TERN-701 inhibited cell proliferation in various clinically relevant mutant CML cell lines (including T315I) with IC 50s ranging from 5 to 26 nM and 70 to 1200 nM against active site and myristoyl site mutations, respectively. TERN-701 was highly selective for BCR::ABL1, with no appreciable activity (>50%) against >450 purified kinase targets. Against a panel of cancer cell lines, TERN-701 was more selective than the comparator allosteric BCR::ABL1 inhibitor asciminib while maintaining similar potency. In vitro combination studies previously revealed that TERN-701 works synergistically with multiple TKIs in the non-mutant K562 cell line. Here, expanded studies using KCL22-s and BCR::ABL1-T315I mutant cell lines identified strong synergistic interactions between TERN-701 and active site TKIs, including ponatinib and dasatinib. Furthermore, prospective mutagenesis studies showed that the combination of TERN-701 and imatinib resulted in greater inhibition of the outgrowth of resistant clones relative to either compound alone.

TERN-701 is a potent and highly selective allosteric inhibitor of BCR::ABL1 that can act synergistically with active site TKIs in vitro, even against the T315I gatekeeper mutation that confers resistance to all approved active site TKIs except ponatinib, which has known safety liabilities. These data support the continued development of TERN-701 for the treatment of CML using both monotherapy and combination approaches.
A Comprehensive Echocardiographic Portrait of Adult Sickle Cell Disease Patients in the Modern Era
Authors: Chan Kok Hoe , Liao Zean , Idowu Modupe
Published: December 2023

Background

Cardiovascular complications, especially pulmonary hypertension, are a known cause of mortality in patients with sickle cell disease (SCD). With advances in medicine and the introduction of various disease-modifying medications, the incidence and prevalence of cardiovascular complications are poorly documented.

Aim

To determine rates of cardiovascular complications in patients with SCD in the modern era of disease-modifying medications in a single center.

Method

We performed a retrospective observational single-center study of patients with SCD: all genotyped and ≥16 years old who had a recent echocardiogram performed during the study period of 2019-2023. Demographics, echocardiogram results, and laboratory findings were collected and analyzed. Descriptive statistics are presented in percentages, medians, and interquartile ranges. An unpaired Student's t-test and Fisher's exact test were used to determine, where appropriate, the level of significant differences among continuous and categorical variables, respectively, with p value <0.05 considered statistically significant. Data was analyzed using GraphPad Prism version 9.0.2.

Results

As of June 2023, 80 patients (38 male and 42 female, median age 30 years) had undergone a routine transthoracic echocardiogram. Sixty-five patients had the HbSS genotype, 8 HbSC, and 7 HbSB null. Sixty-one patients had received at least one treatment, and 19 (24%) were on concurrent therapy. Hydroxyurea was the common SCD-modifying therapy that the patients received (51 patients), followed by L-glutamine and voxelotor (11 each) and crizanlizumab (9 patients).

In total, 56 (70%) patients had at least one cardiac abnormality. The most common cardiac abnormality was left atrial dilatation (41%), followed by left ventricular hypertrophy (33%), right atrial dilation (24%), and left ventricular dilatation (23%). Pulmonary hypertension was present in 30 (38%) patients with tricuspid regurgitation velocity (TRV) ≥2.5 msec (Table 1).

The cohort of patients with pulmonary hypertension tended to be older (median age 31 years, p=0.0357), have a higher creatinine level (median 0.8, p=0.0414), and have coexisting echocardiographic abnormalities such as left atrial dilatation (70% vs. 24%, p<0.0001) or right atrial dilatation (40% vs. 14%, p=0.0136) than SCD patients without pulmonary hypertension (Table 2).

Conclusion

We present a comprehensive echocardiographic assessment of patients with SCD. Despite advances in the treatment of SCD, there remains a high prevalence of cardiac involvement in SCD patients. We found out that 70% of the patients we reviewed had at least one cardiac abnormality on echocardiography. Left atrial dilatation and left ventricular hypertrophy were the most common cardiac abnormalities noted. Pulmonary hypertension was observed in 37.5% of patients with co-existing right and left atrial enlargement. Larger studies are needed to corroborate these findings.
Effect of HLA-DPB1 Mismatch on Incidence and Severity of Chronic Graft-Versus-Host Disease Following Haploidentical Transplantation
Authors: Solomon Scott R. , Solh Melhem M. , Bashey Asad , Bachier-Rodriguez Lizamarie , Zhang Xu , Morris Lawrence , Holland H. Kent
Published: December 2023

Chronic graft-versus-host disease (cGVHD) is an important cause of late morbidity and mortality after haploidentical donor transplantation (HIDT), occurring in up to one-third of patients. The treatment of cGVHD can continue for years and can significantly impact quality-of-life in long-term transplant survivors. Therefore, it is important to identify risk factors that are associated with the incidence and severity of cGVHD. To this end, we investigated 335 consecutive recipients of HIDT utilizing post-transplant cyclophosphamide (PTCy) from 2005-2021, with a particular focus on class II HLA mismatches given prior published associations of HLA-DR and -DP mismatches with HIDT outcomes. HIDT recipients had a median age of 50 (19, 80). PBSC was the graft source in 81%, and conditioning was myeloablative in 49%. Whereas the incidence and severity of cGVHD did not differ between transplants with or without an HLA-DRB1 or -DQB1 allele-level mismatch, the presence of an HLA-DPB1 mismatch was associated with higher 1-yr incidence of all-grade (32% vs. 22%, p=0.07) and moderate-to-severe (22% vs. 12%, p=0.011) cGVHD (see figure). The effect of HLA-DPB1 mismatch on cGVHD was independent of permissiveness by the T cell epitope (TCE) algorithm or the vector of the mismatch (bidirectional, GVH- or HVG-only). In multivariate analysis, an HLA-DPB1 mismatch was associated with an increased risk of moderate-to-severe cGVHD (HR 2.05, p=0.044), while controlling for other covariates (CMV status, year of transplant). Other class II loci (HLA-DRB1 and HLA-DQB1) mismatching were not associated with chronic GVHD incidence in MVA. In summary, the presence of an HLA-DPB1 mismatch is associated with an increased incidence of moderate-to-severe cGVHD following PTCy-based HIDT. This effect, of course, must be balanced by the protective effect of a TCE nonpermissive HLA-DPB1 mismatch on relapse and survival, when selecting the optimal haploidentical donor.
Impact of an HLA-DPB1 Non-Permissive Mismatch By the Revised T Cell Epitope (TCE)-Core Algorithm on Survival Following Haploidentical Donor Transplantation (HIDT) with Post-Transplant Cyclophosphamide (PTCy)
Authors: Solomon Scott R. , Aubrey Michael T , Bachier-Rodriguez Lizamarie , Solh Melhem M. , Jackson Katelin C , Zhang Xu , Roark Christina , Holland H. Kent , Morris Lawrence , Bashey Asad
Published: December 2023

The presence of an HLA-DPB1 non-permissive mismatch (NPMM) by the TCE-3 model has been associated with improved survival following HIDT utilizing post-transplant cyclophosphamide (PTCy). A revised model (TCE-core) which further separates TCE “group 3” alleles into “core”(C) and “non-core” (NC) alleles has been developed (Arrietta-Bolanos et al. Blood 2022). In this new model, a formerly permissive mismatch (PMM) resulting from “group 3” alleles in both donor and recipient are now considered a C-NPMM if one or more of those alleles is NC (see figure), and the total number of NPMMs by the TCE-core model results from adding these C-NPMMs to conventional NPMM. In order to study the impact of an HLA-DPB1 NPMM by the TCE-core model on outcomes following PTCy-based HIDT, we evaluated 242 consecutive HIDT recipients with ALL, AML or MDS transplanted between 2005 - 2021 at a single institution, (median age 51 [19,80], 38% non-White, 32% DRI high/v. high). We further examined the effect of the vector of non-permissiveness (see figure), hypothesizing that a NPMM in the GVH or bidirectional direction may have a more significant impact on relapse and survival than an HVG-only NPMM. Median follow-up was 62 [23, 199] months. A NPMM was seen in 129 donor-recipient pairs (53%), of which 65 (27%) were either in the GVH-only or bidirectional vector. In univariate analysis, a trend for improved 5-yr OS was seen in the context of a TCE-core NPMM vs. matched/PMM HLA-DPB1 (59% vs. 49%, p=0.06). Restricting TCE-core NPMM to the GVH/bidirectional vector further improved 5-yr OS (64% vs. 49%, p=0.023, see figure). Compared to an HVG-only NPMM, a GVH/bidirectional NPMM was associated with a significantly lower 5-yr risk of relapse (23% vs. 45%, p=0.016). In Cox multivariable analysis, adjusted for disease risk index, regimen intensity and donor age, a GVH/bidirectional mismatch was associated with improved OS (HR 0.62, 95% CI 0.38-0.99, p=0.047), compared with HLA-DPB1 match/PMM. Although not statistically significant, a trend was also seen for DFS (HR 0.69, 95%CI 0.44-1.09, p=0.11) and relapse (HR 0.63, 95%CI 0.35-1.14, p=0.12). Compared with an HVG-only TCE-core NPMM, a GVH/bidirectional NPMM was associated with significantly lower relapse (HR 0.48, 95%CI 0.26-0.89, p=0.020). In summary, the presence of TCE-core GVH/bidirectional NPMM is associated with less relapse and significantly improved OS following PTCy-based HIDT. This association should be confirmed in a large registry analyses and potentially incorporated in future donor selection algorithms.
The Molecular Prognostic Scoring System for Normal Karyotype Myelodysplastic Syndromes
Authors: Wang Wei , Zhang Yudi , Yang Wenli , Han Yueyuan , Ge Zheng , Tong Hongyan
Published: December 2023

Purpose: Myelodysplastic syndromes (MDS), is characterized by clinical heterogeneity and varying prognoses. Reliable prognostic models are essential to stratify MDS patients and optimize clinical management. Although incorporating molecular information into MDS prognostic models improves their predictive ability, limitations of next-generation sequencing panels and potential heterogeneity among MDS patients with different karyotypes hinder their clinical translation.

Methods: In total, 237 normal karyotype (NK) MDS patients from the First Affiliated Hospital of Zhejiang University School of Medicine were used as the derivation set to investigate the molecular frequency and prognostic spectrum. A parsimonious molecular-clinical model was developed with overall survival (OS) as the endpoint and compared with other models using the concordance index as the primary evaluation index. An external independent validation set from the International Working Group for Prognosis in MDS cohort(691 NK MDS patients) was used to evaluate the model generalization ability.

Results: In NK MDS, the gene mutation frequency spectrum is similar to the overall MDS population, the differences mainly lie in the prognostic spectrum. Genes such as TP53, previously associated with MDS prognoses have lost their prognostic value. Therefore, a seven-parameter molecular-clinical prognostic model called the Molecular Prognostic Scoring System for NK MDS (NK-PSS-M) was developed based on a specific molecular prognostic spectrum of NK MDS. The model includes age, hemoglobin, platelet count, blast percentage, CEBPA, RUNX1, and U2AF1. The NK-PSS-M model allows for personalized risk assessment based on patient characteristics, with higher scores indicating a greater risk and poorer prognosis. In addition, we developed a five-class classification system based on these scores, which differentiates patients across risk categories and predicts outcomes, such as OS and LFS(Figure 1 A and B). The performance of NK-PSS-M was comparable to that of the Molecular International Prognostic Scoring System(IPSS-M), and it significantly outperformed IPSS-R and other molecular prediction models based on it. The above results were confirmed in the external independent validation cohort(Figure 1C). Our transcriptomics analysis revealed that the low-risk group defined by NK-PSS-M demonstrated significant activation of the inflammatory pathway, and immune activation, which is consistent with the current understanding of the immunological landscape of MDS.

Conclusion: The NK-PSS-M model improved the risk stratification of non-molecular models and provided a reliable alternative to the IPSS-M. In addition, we demonstrated the molecular heterogeneity of MDS across different cytogenetics and emphasized the importance of developing cytogenetics-specific prognostic models.
Long-Term Hematopoietic Dysfunction in Patients with Single Large-Scale Mitochondrial DNA Deletion Syndromes
Authors: Greenberg-Kushnir Noa , Barg Assaf Arie , Schiby Ginette , Mardoukh Corine , Varda-Bloom Nira , Bar Diana , Anikster Yair , Bielorai Bella , Toren Amos , Jacoby Elad
Published: December 2023

Pearson syndrome (PS) is the only single large-scale mitochondrial DNA deletion syndrome (SLSMDs) presenting substantial hematologic manifestations, usually a severe though transient cytopenia of childhood. Recently, progression to myeloid malignancies has been described among PS patients. Older PS patients, as well as patients with SLSMDs presenting with Kearns-Sayre syndrome (KSS) or with progressive external ophthalmoplegia (PEO), do not display hematologic abnormalities. Since the underlying mitochondrial DNA deletion is similar in all SLSMDs, we hypothesized that bone marrow (BM) dysfunction is a common and persistent manifestation in these patients.

We studied 16 patients with molecular-confirmed SLSMDs: 13 with PS, and three with KSS. The median age at diagnosis was 25 months (range 2-136 months). Seventy-five percent of patients presented with cytopenia in infancy, and 56% required blood products. Resolution of cytopenia occurred in 92% of this subgroup, at a median age of 24 months (range 12-39 months).

BM studies were performed in all patients, and 56% of patients had multiple BM evaluations. All BM biopsies examined showed signs of marrow dysfunction, including in patients without a history of cytopenia. Median BM cellularity was 50% (range 30-80%) at a median age of 5 years. BM samples showed a paucity of precursor myeloid cells, signs of dyserythropoiesis, and abnormal or missing megakaryocytes. We report low hematopoietic stem and progenitor cell functionality in patients with SLSMDs: The median CD34+ count was 0.68% (range 0.27-1.92%, expected range 1-2%). BM colony forming unit (CFU) capacity was reduced in SLSMD patients: the median CFU in patients was 280 per 10 6 cells compared to 1090 per 10 6 cells in healthy controls (p=0.004). Conventional cytogenetic analysis was done on 10 patients (62%), and FISH analysis was done on 15 patients (93%). Monosomy 7 was identified in seven patients (44%): in one patient the monosomy 7 resolved in a follow-up BM assay. Another patient with signs of myelodysplasia and 30% of the BM cells having monosomy 7 was referred to an allogeneic bone marrow transplantation.

To conclude, long-term BM dysfunction is evident in all patients with SLSMD and includes a risk of developing clonal evolution and monosomy 7. This data, combined with other recent publications, highlights the ongoing marrow failure and suggest that SLSMD syndromes should be included under inherited BM failure syndromes.