bluuebunny/crossref_metadata_2025 huggingface dataset:
CPT1a Controls the Function of Long-Term Hematopoietic Stem Cells By Regulating the Balance between Fatty Acid Oxidation and Oxphos in Mitochondria
Authors: Hashimoto Michihiro , Li Jue , Andreassen Paul R. , Huang Gang
Published: December 2023

Hematopoietic stem cells (HSCs) rely on the balance in place of quiescence and self-renewal to sustain stem cell potential and differentiation to generate mature blood cells. Appropriate regulation of cellular and mitochondrial metabolic processes such as glycolysis, OXPHOS, fatty acid metabolism, and amino acid metabolism are required for HSC maintenance. However, the mechanisms by which cellular and mitochondrial metabolisms regulate HSC quiescence, proliferation, and differentiation remain to be fully understood. Fatty acid oxidation (FAO) is a metabolic process involving the breakdown of fatty acids into acetyl-CoA, which is then transported into mitochondria by carnitine palmitoyltransferase 1a (CPT1a). It has been reported that inhibiting FAO using non-specific inhibitors of CPT1a impairs HSC maintenance. Interestingly, CPT1a forms a complex with the Fanconi anemia complementation group D2 (FANCD2) protein (T Zhang et al., Scientific Reports, 2017). Given that Fanconi anemia is associated with HSC defects, this CPT1a complex may play a critical role in maintaining fatty acid dependent mitochondrial metabolism that mediates HSC dormancy and stemness. However, the exact role, in HSCs, of mitochondrial CPT1a, the rate-limiting enzyme in FAO, remains unknown. Here, we find that CPT1a is highly enriched in hematopoietic stem cells. Further, we investigated the role of CPT1a in hematopoiesis utilizing a CPT1aconditional knockout in mice driven by Vav1-Cre. The CPT1a knockout ( Cpt1aΔ/Δ) displays profound HSC defects, including loss of HSC self-renewal and accelerated differentiation. Mechanistically, we find that loss of Cpt1a results in increased ATP production, mitochondrial membrane potential (Δφm), and mitochondrial ROS accumulation in long-term HSCs (LT-HSCs), as well as defective LT-HSC function indicated by bone marrow transplantation and 5-FU genotoxic stress assays. Furthermore, utilizing mitochondrial complex formation assays and measurements of mitochondrial complex activity, BM cells of Cpt1aΔ/Δ mice display enhanced mitochondrial complex formation and up-regulation of mitochondrial complex activity. Our results suggest that Cpt1a regulates FAO transport and that loss of Cpt1a deregulates mitochondrial metabolisms including oxidative phosphorylation (OXPHOS). Taken together, our study suggests that CPT1a plays a key role in LT-HSC maintenance by controlling the balance between FAO and OXPHOS in mitochondria. Because of the noncanonical relationship between CPT1a and FANCD2 for the regulation of mitochondrial metabolism, correction of fatty acid metabolism and OXPHOS may present a novel therapeutic avenue for Fanconi anemia.
First Disclosure of AZD9829, a TOP1i-ADC Targeting CD123: Promising Preclinical Activity in AML Models with Minimal Effect on Healthy Progenitors
Authors: Dutta Dipannita , Pan Pan , Fleming Ryan , Andrade-Campos Marcio , Belova Elena , Wheeler Jennifer , Cheung Patricia , Santacroce Paul , Daniels Casey , Sabol Darrin , Jakubison Brad , Recolin Benedicte , Dippel Andrew , Wey Margaret , Shaffer Arthur , Rosfjord Edward , Hurt Elaine , Gorgun Gullu , Younes Anas , Sapra Puja
Published: December 2023

CD123 is a cell surface protein that is overexpressed in several hematologic malignancies including acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) with restricted expression in normal hematopoietic stem cells. Clinical development of an antibody drug conjugate (ADC) with first-in-class opportunity for a Topoisomerase I inhibitor (TOP1i) payload in haem malignancies including CD123-expressing AML and MDS is planned. We describe for the first time the preclinical activity of AZD9829, a CD123-targeting antibody conjugated to Astrazeneca's proprietary TOP1i payload, AZ14170132, with a drug-to-antibody ratio of 8. AZD9829's primary mechanism of action is to deliver TOP1i payload into CD123-expressing cancer cells, leading to DNA damage and apoptosis. Compared to other AML targets like CD33, CD123 is highly expressed in AML patient bone marrow (BM) with limited expression in healthy donor BM (Figure 1). AZD9829 showed robust in vitro killing of CD123-positive AML cell lines and demonstrated targeting of BM-resident tumor cells in AML patients, with mild, transitory effects on the BM compartment from healthy donors. We showed that a single intravenous (IV) dose of AZD9829 was sufficient to induce 100% tumor growth inhibition (TGI) in high and low CD123-expressing AML cell line xenografts at 2 mg/kg and 3 mg/kg, respectively. Anti-tumour activity of AZD9829 was also observed across a panel of 13 AML patient-derived xenograft (PDX) models representing AML disease heterogeneity with diverse mutation status, disease stage, prior treatment response, and expression level of CD123. These disseminated PDX models were treated with weekly IV dosing of AZD9829 at 5 mg/kg, for two doses. AZD9829 achieved ≥50% leukemic blast reduction in blood (7/7 models) (Figure 2) and in bone marrow (6/7 models) at day 14 after the first dose. Furthermore, AZD9829 demonstrated durable blast reduction at day 28 after the first dose with leukemic blast reduction in blood (7/13 models) and in bone marrow (5/13 models). Safety studies in cynomolgus monkey support the clinical development of AZD9829, a promising therapeutic candidate for the treatment of AML across the spectrum of CD123-expression and genetic mutations.
Outcomes of Sarcoidosis on Patients with Sickle Cell Disease - a Review of the National Inpatient Database 2016-2020
Authors: El Sharu Husam , Zweigle Joshua , Jamil Maria , Singh Sukhvir , Cowles Sara , Liles Darla K.
Published: December 2023

Introduction:

Sickle Cell Disease (SCD) is an inherited hemolytic disorder due to an autosomal recessive mutation affecting mainly African Americans. SCD can lead to various complications, including Sickle Cell Crisis (SCC), Acute Chest Syndrome (ACS), strokes, and different osseous complications. Notably, these complications can be provoked using steroids. Sarcoidosis is a multisystem inflammatory disorder that is characterized by the presence of bilateral hilar lymphadenopathy that is commonly treated with steroids. Unfortunately, it is also preferentially affecting African Americans. The complication rates in patients with both diseases have rarely been described. We aim to describe the effect of sarcoidosis on the mortality and complication rates of admitted patients with SCC.

Methods:

Using the National Inpatient Sample (NIS) 2016-2020, we analyzed adult SCC and ACS hospitalizations with and without sarcoidosis using International Classification of Diseases - 10 Clinical Modification (ICD-10-CM) codes. The primary outcome was inpatient mortality. Secondary outcomes were inpatient morbidities, mean length of stay (LOS), and mean total hospital charge (THC). A multivariate logistic regression and linear regression analyses were used to adjust for potential confounders.

Results:

Out of 359655 patients hospitalized with SCC, only 4.6% had concomitant sarcoidosis. Of these, 70% were females, with a mean age of 38.4 compared to an average age of 32 in patients without sarcoidosis (p-value <0.05). The mean LOS for patients with sarcoidosis and SCC was 6.4 days, while it was 5.1 days for patients without sarcoidosis (p-value 0.001). While adjusting for common comorbidities and patients' characteristics, patients with SCC and sarcoidosis did not have a statistical mortality difference compared to patients without sarcoidosis [adjusted odds ratio (aOR): 2.3, CI: 0.4-14). On stratified analysis, patients with ACS and sarcoidosis had increased odds of mortality with an aOR of 6.68, CI: 1.1-39.7, and a p-value of 0.037. Moreover, amongst patients who had SCC, patients with concomitant sarcoidosis had a higher risk of acute coronary syndrome (aOR: 5.1, CI 1.24-21.0) and avascular necrosis (aOR: 1.57, CI: 1.12-2.20). There were no statistically significant differences in the odds of ischemic strokes, transient ischemic attacks, acute and chronic kidney disease, risk of intubation, pulmonary edema, osteoporosis, venous thrombosis, and THC. Figure 1 shows the Forrest plot for multivariate analysis of in-hospital morbidities when adjusted for patient demographics, comorbidities, and hospital characteristics.

Conclusion:

Patients admitted with ACS and sarcoidosis had higher odds of mortality. Patients with SCC and sarcoidosis also had higher odds of acute coronary syndrome, avascular necrosis, and mean LOS. Nevertheless, this has not impacted other inpatient comorbidity.
Effectiveness of Intensive Chemotherapy with 7+3 Versus Vidaza and Venetoclax in Acute Myeloid Leukemia with Sequential Stem Cell Transplant: Results from Real-World Cohorts
Authors: Fuqua Jacob , Abonofal Abdulrahman , Safi Salah Ud Din
Published: December 2023

Introduction:

Intensive chemotherapy(IC) with cytarabine and anthracycline (7&3) has been the backbone therapy for medically fit patients for the last 50 years. The use of Venetoclax and hypomethylating agent combination has be used in older and unfit patient populations. The Venetoclax combination regimen has improved outcomes in the selected group but has been characterized to be inferior to 7&3 regimen among some studies. The role of stem cell transplant has not been previously discussed who receive IC versus Venetoclax based chemotehrapy

Methods:

Retrospective data was extracted using the TriNetX database. The primary endpoint was overall survival, and stem cell transplant in the two cohorts.

Results:

5590 patients were identified in cohort-1 (7+3 cohort) and 2944 patients were identified in cohort-2 (Vidaza with Venetoclax cohort). Table-1 shows patient characteristics at baseline. Propensity-score-matching was used to balance the two cohorts. Patient count in each group after matching was 1393 (Table-2). Overall survival was 43.78% in cohort-1 versus 16.56 % in cohort-2, with HR of 0.510 (95% CI 0.457, 0.569) in favor of cohort 1 (Figure-1) at 5-year mark. Median OS for cohort-1 was 1055 days versus 289 days for cohort-2. Additional analysis was performed to determine number of patients who underwent a Stem Cell Transplant in each cohort and the effect of transplant on outcomes. 462 patients in cohort-1 and 358 patients in cohort-2 underwent transplant respectively. Overall survival was 51.29% in cohort-1 versus 61.64% in cohort-2, with HR of 1.180 (95% CI 1.027, 1.254) in favor of cohort-2 at 5-year mark. Median overall survival was not reached for either cohort at the data cut-off.

Conclusions:

Our retrospective analysis of a large patient cohort showed improved OS and decreased risk of death in patient's undergoing 7&3 induction therapy as compared to Vidaza with Venetoclax. In patients who underwent stem cell transplant, the overall survival was statistically better in Vidaza with Venetoclax cohort as compared to 7&3 induction therapy cohort. To the best of our knowledge, this is the first large database based study evaluating these two induction regimens accounting for stem cell transplantation.
No Improvement in the Age Adjusted Mortality of Older Adults with Acute Lymphoblastic Leukemia Despite the Availability of Blinatumumab, Inotuzumab and Novel TKIs : A 1999-2020 Analysis in the United States
Authors: Varughese Tony , Kabat Maciej , Zolotov Eli , Koprivnikar Jamie
Published: December 2023

Introduction: Acute Lymphoblastic Leukemia (ALL) has a bimodal distribution and while it is the most common malignancy in children, it can also present in adults 55 years of age and older. Adverse disease biology and comorbidities that preclude delivering curative regimens are factors that underlie the markedly disparate outcomes seen between childhood and older adult ALL. Advancements in therapy have improved prognosis in children, however in adults' treatment outcomes are worse. Our study aims to compare mortality trends in older adults compared to children within the United States with ALL.

Methods: Using the Center for Disease Control and Prevention Wide-Ranging Online Data for Epidemiologic Research database, we reviewed death certificate data for ALL patients from 1999 to 2020, age 55 and older in the United States. ICD code C91.0 (Acute lymphoblastic leukemia - Malignant neoplasms) was used to identify patients with ALL. Age-adjusted mortality rates (AAMRs) were calculated per 1,000,000 people and stratified patients by sex, race/ethnicity, and census region (Northeast, Midwest, South, and West). A comparison group between ages 0 and 15 was added with an identical stratification of sub-group data.

Results: In the United States from 1999 to 2020 there were 17,238 deaths in ALL patients 55 and older. For this population there was not an improvement in mortality over the last twenty years (10.8, AAMR 1999) and (10.6, AAMR 2020). Further subgroup analysis did not reveal a significant difference over the years by reported sex, race, and US census region. Men and hispanic patients had the highest AAMR consistently, 12 and 13.6 in 2020. There was a statistically significant improvement in ages 0-15 for overall AAMR, 3.5 in 1999 to 2.2 in 2020 (Table 1).

Conclusion:

There has not been a significant improvement in AAMR for adults 55+ with ALL despite improvements seen in the pediatric patient population. Over the last twenty years AAMR in the older adult population has unfortunately been stagnant. This is despite the approval of several new therapies for adult patients with ALL including blinatumumab, inotuzumab, and the increased recognition of the importance of targeted tyrosine kinase inhibitor-based therapy for patients with Ph+ ALL, which is certainly much more prevalent in the older adult ALL population. The lack of AAMR despite the numerous seemingly therapeutic advances made in the adult population underscores the need to specifically study new therapeutics, their outcomes, utilization, and toxicities, in the sub-population of older adults with ALL in order to understand the lack of collective benefit seen in this unique patient population. Continued monitoring of the AAMR trends in this patient population will soon begin to reflect the availability of CART therapy.
Anti-BCMA Dual Epitope-Binding CAR-Marrow Infiltrating Lymphocytes (MILs) Could Offer a Potent Innovative Immunotherapeutic Tool Against Multiple Myeloma
Authors: Vo Manh-Cuong , Jung Sung-Hoon , Nguyen Van-Tan , Tran Van-Dinh-Huan , Kim Sang-Ki , Bae Woo Kyun , Kim Hyeoung-Joon , Wang Lin , Lee Je-Jung
Published: December 2023

Introduction: Chimeric antigen receptor (CAR) T cells are a promising new treatment for multiple myeloma (MM). Marrow-infiltrating lymphocytes (MILs) derived from MM patient's bone marrow are another potential innovative T cell source for CAR-based cell therapy, capable of robustly and specifically targeting myeloma cells through polyclonal targets. MILs possess polyclonal central memory T cell populations, which could be as effective MM killings via polyantigenic cytotoxic T cells with greater recognitions of broad range of tumor antigens via the polyclonal memory TCR as well as the CAR pathway with strong recognition of tumor antigens. Nanobodies are currently the smallest stable antibody molecules having high affinity for antigen, allowing them to strongly penetrate tumor tissue and even directly to the blood-brain barrier while remaining low toxicity and immunogenicity. With the peculiar properties, nanobodies have been widely used as the antigen biding domain of CAR-T cell. In this study, we investigated anti-B-cell maturation antigen (BCMA) dual epitope-binding CAR-MILs against MM.

Methods: CD3 + T cells isolated from bone marrow mononuclear cells of MM patients were stimulated with Ex Vivo CD3/CD28 in the presence of recombinant human IL-2. The nanobodies VHH01 and VHH02 were provided as dual epitope-binding for BCMA (Senlang, China). After expansion and activation for 2 days, anti-BCMA dual epitope-binding CAR was transduced to the activated MILs and, they were stimulated with Ex Vivo CD3/CD28 in the presence of recombinant human IL-2, IL-7, IL-15, and IL-21. Expanded and activated BCMA dual epitope-binding CAR-MILs (BCMA CAR-MILs) were harvested on days 7, 14, and 21 for further experiments.

Results: The transduction efficiency of BCMA CAR-MILs was approximately 34%, and the expansion of BCMA CAR-MILs was > 100 folds over 21-day culture period. Additionally, BCMA CAR-MILs showed a dramatically increased proportion of CD8 + T cells, while the proportion of CD4 + T cells decreased. Moreover, BCMA CAR-MILs were enriched with a higher proportion of central memory T cells (Tcm) (80%), while the proportions of regulatory T cells (Tregs) and checkpoint molecules (TIGIT, TIM3, and CD73) were not so high. The BCMA CAR-MILs showed potent cytotoxicity, particularly high levels of CD107a degranulation against BCMA-expressing target cancer cell lines, such as Daudi, Raji, RPMI8226, and U266B1 cells, and autologous CD138 + primary myeloma cells.

Conclusion: This study suggested that BCMA dual epitope-binding CAR-MILs were easily expanded and activated using bone marrow mononuclear cells from MM patients with high transduction efficiency and induced a potent anti-myeloma immune response, which may offer an alternative innovative immunotherapeutic approach against MM patients in the future.
Letermovir Prophylaxis for Cytomegalovirus Reactivation in β Thalassemia Major Children after Hematopoietic Cell Transplantation
Authors: Shi Lingling , Liu Rongrong , Zhang Zhongming , Huang Cuifeng , Liu Lianjin , Yao Yibin , Wei Zhenbin , Wei Shuyi , Gan Zhaoping , Sun Na , Liu Beicai , Huang Rui , Zhou Xuemei , Chen Yinghua , Liu Qifa , Lai Yongrong
Published: December 2023

Background: Cytomegalovirus(CMV) is one of the most important infections following allogeneic haematopoietic stem cell transplantation (HSCT). Although Letermovir is licensed for prophylaxis of CMV infection in allogeneic hematopoietic cell transplant adult CMV seropositive patients. There are limited data for letermovir as primary CMV prophylaxis in pediatric patients . The objective of the study was to analyze the the use of letermovir for prophylaxis from CMV infection in β thalassemia major children undergoing allo-HSCT in a single center.

M ethods : This study was an retrospective, single-center evaluation of β thalassemia major pediatric recipients treated by allo-HSCT in the First Affiliated Hospital of Guangxi Medical University. 34 patients included in the study were CMV-seropositive, underwent allo-HSCT, and were started on letermovir for primary CMV prophylaxis between June 2022 and July 2023. CMV monitoring was performed 2 times a week until day +100 after HSCT. The primary end point was CMV reactivation on days +100.

Results: 34 patients were analyzed. 20 patients were transplanted from haploidentical donor, 9 patients were transplanted from matched-related donor, 5 patients were transplanted from matched-unrelated donor. The median followed-up was 129 days (range, 50-405 days).The median age was 9 years (range, 2-16 years). The GVHD prophylaxis for matched-related donor group was cyclosporine-based , and the GVHD prophylaxisfor haploidentical and matched-unrelated donor groups were tacrolimus-based. Letermovir start at median 21 days (range 10-35 days) after HSCT. The median weight at the initiation of letermovir was 18.8 kg (range, 12-25 kg). The dose of letermovir was 240mg and 120mg orally once daily in tacrolimus co-administration and cyclosporine co-administration, respectively. During the follow-up period , CMV reactivation was observed in 8 patients totally (23.5%). Among them, 6 patients(17.6%) were in haploidentical donor group. 1 patient (2.9%) in matched-related group and 1 patient (2.9%) in matched-unrelated donor group was CMV reactivated, respectively. Throughout letermovir prophylaxis, the median CMV DNAemia level was 1945 copies (range,526-3800 copies), and the CMV DNAemia lasting time were 17.5 days (range,15-59 days). No patients developed clinically significant CMV infection. 1 patient died because of pulmonary infection of mucor. No serious hepatotoxicity, nephrotoxicity or letermovir intolerance was described.

Conclusion: Our data support letermovir prophylaxis efficacy and safety in β thalassemia major children after allo-HSCT.
Modeling Gilteritinib Resistance By Enforced cxcl8a Expression in Zebrafish HSPCs
Authors: Oyman Konur , Calkins Donn Lee , Li Wantong , Buelow Daelynn , Baker Sharyn D. , Blaser Bradley Wayne
Published: December 2023

In Acute Myeloid Leukemia (AML), the FMS-like tyrosine kinase III Internal Tandem Duplication (FLT3-ITD) is common and increases the risk of relapse which is only partially mitigated by tyrosine kinase inhibitor (TKI) therapy. Gilteritinib is the current mainstay for treating relapsed FLT3-ITD AML, but therapy is non-curative, and the duration of remission is transient. Mutations in the RAS pathway are known to promote gilteritinib resistance, however non-mutational mechanisms of resistance remain under-explored and could provide new avenues for combination therapy. Our previous work has associated AML cell-autonomous expression of chemokines such as CXCL8 (IL-8) as a mechanism of resistance to gilteritinib therapy. Here we enforce expression of cxcl8a ( cxcl8, ENSDARG00000104795) by hematopoietic stem and progenitor cells (HSPCs) in transgenic zebrafish embryos using a defined Runx1 enhancer element. In the absence of gilteritinib, Runx1:cxcl8(+) animals had greater numbers of HPSCs compared to Runx1:cxcl8(-) animals (4.0 vs 2.1, p = 0.04). When treated with gilteritinib at 100 nM from 48-72 hours post-fertilization (hpf), HSPC numbers were decreased in both groups; however, Runx1:cxcl8(+) animals still had more HSPCs compared to Runx1:cxcl8(-) animals (1.6 vs 0.4, p = 0.02). This suggests that cxcl8 may act as a resistance factor for gilteritinib in vivo and that understanding regulators of cxcl8 expression may provide insight into gilteritinib resistance. Reanalysis of published single cell RNA-seq data in AML patients revealed an association between expression of CXCL8 and BTK and BMX but not other Tec-family kinases such as ITK and TEC itself. Zebrafish do not have an annotated BMX gene; BTK is expressed in HSPCs and neutrophils but not the erythroid lineage. CHMFL is a specific and potent inhibitor of human BMX with an unknown spectrum of activity in zebrafish. In Runx1:cxcl8(-) Runx1:GFP zebrafish, CHMFL treatment reduced HSPC numbers, but this was not rescued in Runx1:cxcl8(+) animals (untreated vs treated, cxcl8(-): 2.9 vs 1.5, p = 0.02, untreated vs treated, cxcl8(+): 4.0 vs 1.4, p = 0.0006). This contrasts with our prior work with the non-specific Tec family kinase inhibitor, ibrutinib, where HSPC numbers were rescued in Runx1:cxcl8(+) animals. Ongoing studies are focused on understanding the targets of ibrutinib and CHMFL in the zebrafish and how these pathways may affect gilteritinib resistance.
Preclinical Efficacy of a Novel CD8-Targeted Fusosome for In Vivo CAR T Cell Therapy
Authors: Chaturvedi Vandana , Green Jesse , Zipp Garrett , Lampano Aaron , Liang Ouwen , O'Rourke Allison , Granger Brian , Elpek Kutlu G , van Hoeven Neal , Fry Terry
Published: December 2023

SG299 is a CD8-targeted fusosome; a self-inactivating (SIN) lentiviral vector (LVV) pseudotyped with a CD8-targeted fusogen, which carries a transgene encoding a CD19-directed chimeric antigen receptor (CAR). We evaluated the preclinical efficacy of a representative CD8/CD19CAR fusosome comparable to SG299 for its ability to control tumor growth in vivo in an NSG mouse model engrafted with human CD19 positive tumor cells and human peripheral blood mononuclear cells (PBMC). We compared direct intravenous (IV) administration and ex vivo exposure of fusosome to resting T cells followed by IV injection of vector-cell mixtures, as has been proposed for extracorporeal delivery (ECD) methods. We have also evaluated the performance of multiple lots of fusosomes to assess consistency in manufacturing and have evaluated donor-to-donor variability in five PBMC donors in this model.

To measure anti-tumor efficacy, immunodeficient NSG mice were challenged IV with NALM6 tumors expressing firefly luciferase (5E5 cells/mouse) on day 4. For direct IV, PBMC (1E7 cells/mouse) were injected IV on day -1 followed by CD8/CD19CAR fusosome (1E7 IU/mouse) on day 0. For ECD, PBMC were incubated with CD8/CD19CAR fusosome for 1 hr and the cell and fusosome mix were injected IV on day 1. Tumor growth was monitored by bioluminescence imaging over a 28-day period. In addition, we have investigated both pharmacokinetics of fusosome induced CAR T cells and NALM6 tumors by flow cytometry.

CD8/CD19CAR fusosome demonstrated statistically significant control of NALM6 tumors by both direct IV and ECD routes of delivery. Kinetic analysis of peripheral blood demonstrated tumor control and in vivo CART cell generation were dose dependent. The peak CAR T cell response was at D14. CARs detected at all timepoints were CD8+ specific; no CD4 + CAR T cells were observed. In addition, all mice that received fusosome demonstrated protection from morbidity and mortality as measured by reduced weight loss and improved survival compared to control or tumor only mice. Lot-to-lot performance variability of fusosomes was minimal; three lots of fusosome that had similar physical and functional attributes showed efficient and equivalent tumor control with multiple PBMC donors. Finally, an assessment of donor-to-donor variability across five PBMC donors using a single lot of fusosome showed donor-specific variations in tumor control as measured by BLI but demonstrated a consistent protection from morbidity and mortality. Collectively, these studies demonstrate preclinical efficacy of our CD8/CD19CAR candidate, lot consistency and verify our ability to generate CD19-directed CAR T cells in vivo using a specifically targeted platform.
Clinical Characterization of Patients with Venous Thromboembolic Disease in 2 Reference Centers in El Salvador
Authors: Rivas Luis Fernando
Published: December 2023

Introduction. During the last decade, venous thromboembolic disease (VTE) has become increasingly important due to its high impact on morbidity and mortality in patients. Suspicion scores together with the diversification of diagnostic methods have allowed rapid and timely diagnosis. Although the annual incidence of VTE is estimated to be approximately 1 in 1,000 people, the risk in certain groups is significantly higher. For the United States, the statistics do not vary significantly. It is estimated that DVT and PE affect between 350,000 and 600,000 people a year, causing between 100,000 and 300,000 deaths. For El Salvador, there are no clinical data related to risk and development factors. Aim. To describe the clinical characteristics of patients with venous thromboembolic disease in 2 reference centers of the Salvadoran Social Security Institute in 2020. Methods. This study is retrospective, observational, cross-sectional and descriptive. All patients with a first-time or recurrent diagnosis that were documented by venous Doppler of the lower and/or upper limbs, computed tomography angiography, magnetic resonance imaging and/or venography at the General Hospital, Medical Surgical and Oncological Hospital of Social Security were included. diagnosed in the internal medicine hospitalization area and in outpatient consultation for the first time in the Hematology Service during the period established between January and December 2020. Results. Of the total of 138 registered patients, the final study population that met all the inclusion criteria was 88, of which the majority corresponded to the female gender in 56.8% (n=50), with a mean age for all of them. the cases of 55 years (range 23 to 88 years). Regarding the associated comorbidities, the 2 main ones were arterial hypertension 52.3% (n=46) and diabetes mellitus 25% (n=22), the most prevalent risk factor being the use of OACs with 9.1% (n=8 ). The most frequent anatomical location was the left lower limb with 45.2% (n=46) of the cases and at the time of diagnosis the initial treatment of all patients with thrombosis was with low molecular weight heparins in 51.1% (n=45). ). VTE in any of its entities continues to be a disease predominantly in adults and women, which is favored by various modifiable and sometimes preventable risk factors. Conclusions. VTE in any of its entities continues to be a disease predominantly in adults and women, which is favored by various modifiable and sometimes preventable risk factors. The main anatomical location of thrombosis is in the lower limbs, mainly on the left side, and in the lungs in main branches. Regardless of the location, mortality is similar in deep vein thrombosis and pulmonary embolism.
Association of Autoimmune Diseases and Their Effect on Outcomes of Heparin-Induced Thrombocytopenia: An Analysis of the National Inpatient Sample (2016-2020)
Authors: Srivastava Arnav , Jain Tripti , Mahmoud Amir , Kharel Himal , Orakzai Abdullah , Eltaher Basant , Piplani Shobhit
Published: December 2023

Introduction: Heparin-Induced Thrombocytopenia (HIT) is a prothrombotic reaction to heparin exposure caused by antibodies directed towards heparin-platelet protein platelet factor 4 complexes. This thrombocytopenia is associated with and can lead to fatal venous and arterial thrombosis. This study aims to compare outcomes and assess the association of autoimmune diseases with HIT.

Methods: A query of the National Inpatient Sample (2016-2020) was performed for index admissions with a principal diagnosis of HIT and concurrent autoimmune diseases using ICD 10 codes. Patient demographics and outcomes were identified. Univariate analysis with t-test or chi-squared was performed where appropriate. A multivariate regression analysis was used to generate adjusted odds ratios (AOR), adjusting for age, race, sex, and Charlson Comorbidity Index (CCI) score. The P-value significance was set at <0.05. All analysis was performed in STATA MP version 14.2.

Results: A total of 72,935 patients with HIT met the inclusion criteria, of which, 2.88% had an underlying autoimmune disease. Those with auto-immune disease had a greater proportion of Caucasians, females, and younger patients. While there were no statistically significant differences in mortality, embolic, or bleeding events, patients with autoimmune disease were 43% less likely to become septic (AOR 0.57, p-value 0.006). Additionally, patients with autoimmune diseases had a lower mean hospital services charge and length of stay (p<0.001).

Conclusion: Patients with HIT and autoimmune disease showed better outcomes in terms of sepsis, mean length of stay, and hospital services charge without any significant difference in embolic or bleeding outcomes. Underlying autoimmune disease likely does not confer any additional risk to HIT hospitalizations. This might be speculated to be due to the different underlying pathology driving HIT or accompanying immunosuppressants treatment in autoimmune diseases hindering the aggressiveness of immune response.
Safety and Efficacy of Venetoclax Salvage Therapy in Relapsed/Refractory Multiple Myeloma with and without t(11;14)
Authors: Steinhardt Maximilian J , Bittrich Max , Zhou Xiang , Noderer Julia , Riedhammer Christine , Xiao Xianghui , Gawlas Sophia , Weis Philipp , Eisele Florian , Munawar Umair , Han Seungbin , Mersi Julia , Waldschmidt Johannes M , Einsele Hermann , Rasche Leo , Kortüm K. Martin
Published: December 2023

Background:

The BCL2 inhibitor Venetoclax (Ven) is an effective treatment option in multiple myeloma (MM). However, despite the recent inclusion in European MM treatment guidelines, no approval has yet been granted for the treatment of MM and the optimal dosage, combination partners and timing of treatment remain under investigation.

Methods:

We conducted an analysis of 40 MM patients treated with Ven at our institution from November 2017 until June 2022. 64% of them had a t(11;14), all were heavily pretreated with a median of 7 lines of prior lines of therapy, and 75% were penta-refractory. High-risk features were enriched in the cohort (47% for all 38 with FISH available via IMWG criteria). Risk was evenly distributed between patients with and without t(11;14). Patients received either Ven alone or in combination with other MM drugs over a median of 5 cycles.

Results:

The full cohort overall response rate reached 53%, ten patients (25%) achieved partial remission (PR), seven patients (17.5%) went into VGPR and four patients (10%) had a complete remission (CR). Moreover, stable disease (SD) for three months or longer was achieved in seven patients (18%). The median progression-free survival was 5.1 months and the duration of response reached 9.6 months.

Subgroup analysis revealed that patients harboring t(11;14) had significantly increased PFS and OS: Median PFS reached 11.2 vs. 4.4 months and median OS was not reached vs. 12.8 months, respectively.

Overall, hematological AEs grade 3 or higher were documented in 65% of patients and three high-grade infectious complications (7.5%) occurred. Two of these patients died from sepsis, another recovered from pneumonia with severe sequelae. Interestingly, in contradiction to the BELLINI experience, all high-grade infectious complications occurred in the t(11;14) positive cohort.

Conclusion:

In our real-world analysis, Ven demonstrated to be an effective MM treatment option for t(11;14) positive relapsed/refractory MM. However, the significant infectious complications suggest antibiotic prophylaxis and careful monitoring of patients undergoing Ven treatment.
Caspase 8 Deletion Causes Infection/Inflammation-Induced MDS-like Bone Marrow Failure in Mice
Authors: Joshi Kanak , Liu Shanhui , Zhang Lei , Mack Ryan Michael , Runde Austin , Hagen Patrick , Barton Kevin , Breslin Peter , Kini Ameet R. , Zhang Jiwang
Published: December 2023

Myelodysplastic syndromes (MDS) are a heterogeneous group of pre-leukemic hematopoietic disorders characterized by persistent cytopenia in peripheral blood (PB) and normo- or hypercellular and morphologic dysplasia in bone marrow (BM) due to ineffective hematopoiesis, as well as resulting in an elevated risk for leukemic transformation. Bacterial or viral infections increase the severity of MDS patients' symptoms. An inflammatory BM micro-environment and programmed cell death of hematopoietic stem/progenitor cells (HSPCs) have been thought to be the major causes leading to ineffective hematopoiesis. Three types of programed cell death, apoptosis, necroptosis and pyroptosis, were observed in BM tissues of MDS patients, suggesting PANoptosis might play a critical role in the pathogenesis of MDS. Caspase 8 (Casp8) is a master regulator of PANoptosis. Casp8 is downregulated in HSPCs from patients with MDS, and is associated with increased Ripk1 protein, which is another master regulator of PANoptosis. To study the role of PANoptosis in normal hematopoiesis and the pathogenesis of MDS, we generated Mx1-Cre and Rosa-ERT 2Cre-mediated inducible C asp8 knockout mice ( Casp8 -/-). Mx1-CreCasp8 -/- mice died of BM failure within 10 days of three polyI:C injections (administered every other day) due to the massive depletion of HSPCs. This could be completely rescued by Ripk3 deletion, suggesting that HSPCs were eliminated by necroptosis. Rosa-ERT 2Cre- Casp8 -/- mice are completely normal without any significant changes in BM hematopoiesis after induction by five doses of tamoxifen (TAM) when maintained in specific pathogen-free facilities. However, such mice developed a spontaneous BM failure phenotype when maintained in quarantine facilities. This BM failure can be attenuated by antibiotic treatment, suggesting that the phenotype is infection-induced. In addition, low dose polyI:C injections or treatment with LPS also induces BM failure in Rosa-ERT 2Cre - Casp8 -/- mice living in germ-free conditions. Detailed analysis suggested that Casp8 -/- HSPCs are hypersensitive to polyI:C, LPS, TNFα and IFNα/γ-induced cell death. Most importantly, all these phenotypic and functional abnormalities observed in Rosa-ERT 2Cre- Casp8 -/- mice can be completely prevented by Ripk3 deletion. In addition, serial transplantationstudy demonstrated that Casp8 -/- HSCs have impaired self-renewal capacity. Interestingly, the self-renewal of Casp8 -/-Ripk3 -/- HSCs is significantly enhanced compared to Wild-type HSCs. Our study suggests an essential role for a balance in Casp8 and Ripk3 activities in the regulation of normal hematopoiesis and the self-renewal of HSCs, the disruption of which might induce inflammation and BM failure, resulting in MDS-like disease.
Real-World Treatment Patterns Following Update to National Comprehensive Cancer Network Guidelines for Light-Chain Amyloidosis: Results from a Us Administrative Claims Database
Authors: Thompson Jeffrey , Wechalekar Ashutosh D.
Published: December 2023

Background: Light-chain amyloidosis (AL) is a rare, multi-organ disease that can cause serious organ damage. In January 2021, a new therapy, daratumumab, was FDA-approved for the treatment of AL in combination with current standard-of-care (SoC) of bortezomib/cyclophosphamide/dexamethasone (CyBorD) and was classified as SoC according to the National Comprehensive Cancer Network (NCCN) guidelines. The objective of this study is to describe current real-world treatment patterns amongst patients with AL who newly initiated treatment January/2021 through June/2022.

Methods: This study included patients aged ≥18 identified in the IQVIA PharMetrics database (>72 million lives 2021-2022 total) who were newly initiating pharmacological treatment with bendamustine, bortezomib, carfilzomib, cyclophosphamide, daratumumab, ixazomib, lenalidomide, melphalan, pomalidomide, or venetoclax 01Jan2021-30Jun2022 (date of first therapy classified as “index date”). Patients were required to have an ICD-10 diagnosis of AL (E85.81) identified during full study period (01Jan2020-30Sep2022) as well as continuous medical and prescription insurance enrollment from 1 year pre-treatment-index through 90-days post-treatment-index. Patients were excluded if they had any of the following: a claim indicating treatment with stem cell transplant; a diagnosis of familial mediterranean fever; a diagnosis of secondary, transthyretin, or hereditary amyloidosis; a diagnosis of a non-myeloma/non-lymphoma malignancy. Treatment patterns assessed included index treatment regimen and daratumumab adoption/use patterns at index, 30-days post-index, and 90-days post-index.

Results: A total of 194 patients met all eligibility criteria, out of which the majority were male (N=119; 61%), with the largest insurance coverage being self-insured (N=80; 41%), and a mean age of 62 years (SD=9.8). The most common index treatment regimen was CyBorD+daratumumab (N=53; 27%) followed by dexamethasone+daratumumab (N=24;12%) and daratumumab monotherapy (N=16; 8%) ( Table). Across all treatment regimens at index date, daratumumab was found to be utilized by 62% (N=121) of patients. Furthermore, 76% (N=148) and 80% (N=155) of patients were found to have utilized daratumumab within 30-days or 90-days of treatment initiation, respectively.

Discussion: AL is life threatening disease that impacts organ function and quality of life. Until 2021, there were no FDA approved therapies for AL. Daratumumab use was rapidly adopted to treat patients with AL following approval, in accordance with national guidelines, primarily in combination with other treatments such as CyBorD. This has changed the treatment paradigm providing hope to patients and clinicians to treat this devasting disease and should be informative for current and future clinical trial considerations in development of medications to treat AL.
Estrogens and Prolactin Do Not Regulate Maternal and Embryo Iron Homeostasis during Pregnancy
Authors: Sangkhae Veena , Zhang Vida , Ganz Tomas , Nemeth Elizabeta
Published: December 2023

Iron is essential for maternal and fetal health but the molecular mechanisms ensuring increased iron availability during pregnancy are not well understood. Hepcidin is the key iron-regulatory hormone and functions by blocking iron absorption, recycling and mobilization from stores, effectively decreasing plasma iron levels. In healthy human and rodent pregnancies, maternal hepcidin decreases starting in the second trimester and is nearly undetectable by late pregnancy. This decrease in hepcidin allows for greater iron availability for transfer to the fetus. If hepcidin is not appropriately suppressed during pregnancy, it results in fetal anemia, intrauterine growth restriction, and even fetal demise in severe cases 1.

Unrelated to pregnancy, hormones estrogen and prolactin have been previously implicated in hepcidin regulation 2,3, although with conflicting results on the direction of hepcidin change 4-6. The levels of both hormones increase dramatically over the course of pregnancy, thus we hypothesized that they may modulate iron homeostasis during pregnancy. Studying the role of estrogens and prolactin during pregnancy is challenging as both hormones are required for pregnancy. Additionally, mice with global knockout (KO) of either estrogen receptor alpha or prolactin receptor are infertile. Thus, to determine the contribution of estrogens and prolactin on maternal hepcidin suppression during pregnancy, we generated liver-specific knockouts of either estrogen receptor alpha ( Er- α f/f;Alb-Cre +) or prolactin receptor ( Prlr f/f;Alb-Cre +), here abbreviated as ER-α KO and PRLR KO. Female ER-α KO, PRLR KO and WT mice were set up for timed pregnancy, and liver hepcidin mRNA, as well as maternal and embryo iron and hematological parameters were measured at E18.5 . Hepcidin suppression in ER-α KO and PRLR KO dams was not different from that in WT dams, suggesting that estrogens and prolactin are not major mediators of maternal hepcidin suppression. Additionally, maternal and embryo hematological parameters and liver iron concentrations were similar between WT, ER-α KO, PRLR KO pregnancies, further supporting our observation that estrogens and prolactin do not regulate iron homeostasis in pregnancy. The mechanisms by which maternal hepcidin is suppressed during pregnancy remain to be determined.

1Sangkhae V et al, Blood, 2020

2Yang Q et al, Endocrinology, 2012

3Wang J et al, Nanomedicine: nanotechnology, biology and medicine, 2015

4Zhen et al, Hepatology, 2013

5Ikeda Y et al, PloS one, 2012

6Lehtihet M et al, PloS one, 2016
Sickle Cell Disease Electronic Health Record Population Management Approach to Support Delivery of High-Quality Care: Leaving Paper Lists behind
Authors: Smith-Whitley Kim , Brooks Elizabeth , Brandemarte Alicia , Norris Cynthia
Published: December 2023

Children and young adults with sickle cell disease (SCD) require complex health care services to screen for early signs of disease-related complications and to initiate and monitor disease-modifying therapies. Care coordination is challenging as it involves multiple team members across multiple subspecialties. Evidence-based clinical protocols and quality metrics are essential for SCD programs to deliver high-quality care. Systems to track this care delivery are crucial to ensure equitable, accessible care is consistently provided.

In 2016, the Comprehensive Sickle Cell Center (CSCC) and the Population Health Innovation Team at Children's Hospital of Philadelphia (CHOP) developed and implemented an electronic health record (EHR)-based system, the SCD patient registry and proactive care outreach tools, to facilitate processes needed to ensure patient service delivery in a timely fashion. We selected Transcranial Doppler (TCD) ultrasounds, hydroxyurea therapy initiation, and annual clinic visits to monitor stroke risk screening, disease-modification therapy, and general chronic illness care. Between January 2015 and June 2023, approximately 1400 patients with SCD were tracked across two CHOP outpatient clinics, the CHOP Philadelphia and CHOP Voorhees campuses. We tracked TCD completion, hydroxyurea prescriptions, and clinic visits per year for age-eligible patients with SS and SB0thal.

Prior to implementation of the EHR-based tools and new workflows in October 2016, for age-eligible patients, on average 75% of patients had received annual TCD screening, 55% of patients had received hydroxyurea prescriptions within the prior 6 months, and 85% had attended outpatient hematology clinic at least twice within the year. As of June 2023, on average, completed TCD visits had increased to 91%, patients prescribed hydroxyurea had increased to 78%, and patients attending 2 clinic visits within a year had increased to 89%. These metrics remained consistent and improvement was sustained despite the COVID-19 pandemic. Table 1 describes the drivers influencing these improvements, which included various interventions involving social work, nursing, community health workers, registrars, care coordinators, and physicians. Figure 1 illustrates the changes achieved over time in rates of TCD completion, hydroxyurea prescriptions, and clinic visit attendance.

This implementation of an SCD EHR-based population health tool and new care team workflows facilitated more efficient processes for various care team members and improved our ability to consistently provide timely services to children and young adults with SCD. This resulted in sustained improvement in all metrics and high-quality care for our patients.
Late Effects of Allogeneic Stem Cell Transplantation for Non-Hodgkin Lymphoma. Swiss Cohort Analysis 1997-2021
Authors: Rebmann Ekaterina , von Werdt Alexander , Künzi Arnaud , Wolfensberger Nathan , Baldomero Helen , Medinger Michael , Passweg Jakob R. , Nair Gayathri , Masouridi-Levrat Stavroula , Chalandon Yves , Rovó Alicia
Published: December 2023

Background: despite rapid development of new highly efficient targeted and cellular-therapies, allogeneic stem cell transplantation (allo-HSCT) represents so far the only potentially curable treatment according to a longest follow-up in a setting of relapse/refractory Non-Hodgkin lymphoma (rrNHL). However, late effect (LE) issues of allo-HSCT in rrNHL seem to be underexplored. Here, we present the first analysis of a retrospective, multicenter, registry-based analysis (Swiss Blood Stem Cell Transplantation and Cellular Therapy, SBST) of LE in patients who underwent allo-HSCT for rrNHL in Switzerland.

Methods: We retrospectively collected data from 109 patients with all types of rrNHL who underwent allo-HSCT in 3 University Hospitals of Switzerland (Zurich, Basel and Geneva) between May 1997 and November 2021. The inclusion criteria was 2 years survival after allo-HSCT. The primary endpoint was the cumulative incidence (CI) of LE and secondary endpoint were overall survival (OS), relapse incidence (RI) and type of LE according to organ/systems involved.

Results: median of follow-up was 6.5 years. Patients who survived 2 years after allo-HSCT presented 5 and 10 years OS of 94% [95% CI 86.3 - 97.5] and 86% [95% CI 75 - 92.4] respectively. RI and CI of LE were 18.3% and 44% at 5 years and 34.4% and 58% at 10 years post allo-HSCT. A total of 79/109 (72%) of patients experienced any type of LE during the follow-up period. Renal dysfunction (21%), secondary malignancies (18%), cardiovascular disease (18%), osteoporosis (18%) and thyroid dysfunction (17%) were the most frequently observed. CI of secondary malignancies was 6.1% and 20% at 5 and 10 years respectively.

Conclusions: Our analysis of a mixed-type rrNHL cohort confirms the efficacy of allo-HSCT in heavily pretreated/refractory patients, showing excellent OS at 5 and 10 years for those who survived 2 years. Both RI and LE present a competitive risk for survival benefit beyond 2 years after allo-HSCT. Secondary malignancies, renal and cardiovascular disease appear to be the most frequent types of LE, revealing the toxicity accumulated prior to allo-HSCT in the setting of rrNHL. These results should be interpreted with caution due to a retrospective nature of the analysis, the heterogeneity of the conditioning regimen and the NHL cohort.

Figure 1. a) Overall survival (KM-estimate) b) cumulative incidence of late effects