Donor Bone Marrow Research Articles

Preclinical ex vivo IL2RG gene therapy using autologous hematopoietic stem cells as an effective and safe treatment for X-linked severe combined immunodeficiency disease

X-linked severe combined immunodeficiency disease (X-SCID) is a rare inherited disease caused by mutations in the interleukin 2 receptor subunit gamma gene (IL2RG), which encodes the common γ chain protein, a subunit of the receptor for lymphocytes. X-SCID is characterized by profound defects in T-cell, B-cell, and natural killer cell function. Here, we report a Chinese cohort of nine X-SCID patients with six novel IL2RG mutations. Among those, the two adolescent patients carrying missense mutation with an atypical immunotype were confirmed by further analyzing IL-2-JAK-STAT5 signaling, T cell proliferation, and T cell receptor excision circles (Trecs). Interestingly, Bacillus Calmette-Guérin (BCG) disease occurred commonly in this cohort. Although allogeneic hematopoietic stem-cell transplantation is curative for the disease, it is not available to all patients due to the lack of suitable matched donors. Autologous gene therapy using a self-inactivating lentiviral vector (SIN-LV) technology, developed in recent decades has provided an alternative therapy for treating such mono-genetic diseases. Here, we performed the pre-clinical studies to assess our SIN-LV carrying IL2RG on human ED7R cells deficient in IL2RG and CD34+ stem cells derived from the bone marrow of a healthy donor and a patient with X-SCID. This work is done complied with the established "Good Manufacturing Practice" (GMP) used in the clinical trials. In addition, a safety study is performed using the transduced CD34+ cells implanted into the axilla of nude mice in vivo. Overall, our studies have demonstrated the efficiency and safety of SIN-IL2RG-LV, which paves the way for conducting X-SCID gene therapy clinical trials in China in the near future.

A Novel HLA-DPA1*02 Variant, HLA-DPA1*02:141, Identified in a Bone Marrow Donor Candidate From Brazil.

Identification of the novel HLA-DPA1*02:141 allele that differs from HLA-DPA1*02:02:02:01 at one position in exon 4.

Development of iPSC-derived human bone marrow organoid for autonomous hematopoiesis and patient-derived HSPC engraftment

Current efforts in translational studies in hematology often rely on immunodeficient mouse models for engrafting patient-derived hematopoietic stem and progenitor cells (HSPCs), yet these models often face challenges in effectively engrafting cells from patients with various diseases, such as myelodysplastic syndromes (MDS). In this study, we developed an induced pluripotent stem cell (iPSC)-derived human bone marrow organoid model that closely replicates the bone marrow microenvironment, facilitating the engraftment of MDS patient-derived HSPCs, thereby mirroring the patients' distinct disease characteristics. Specifically, through advanced microscopy, we verified the development of a complex three-dimensional network of endothelial, stromal, and hematopoietic cells within the organoids, resembling the autonomous human marrow microenvironment. Furthermore, we showed that HSPCs derived from the donor bone marrow of normal individuals or patients with MDS can migrate to and proliferate within the organoid’s vascular niche while maintaining self-renewal and original genetic profiles. Within the organoids, the differentiation patterns from MDS HSPCs were significantly distinct compared to the multilineage hematopoiesis from normal HSPCs, which can be correlated with the clinical manifestations of the disease. These findings underscore the significance of the organoid model in studying human hematopoiesis and the pathophysiology of hematologic diseases, offering new avenues for personalized medicine and therapeutic interventions.

Multiple measures for self-identification improve matching donors with patients in unrelated hematopoietic stem cell transplant

BackgroundQuestions persist around whether and how to use race or geographic ancestry in biomedical research and medicine, but these forms of self-identification serve as a critical tool to inform matching algorithms for human leukocyte antigen (HLA) of varying levels of resolution for unrelated hematopoietic stem cell transplant in large donor registries.MethodsHere, we examined multiple self-reported measures of race and ancestry from a survey of a cohort of over 100,000 U.S. volunteer bone marrow donors alongside their high-resolution HLA genotype data.ResultsWe find that these self-report measures are often non-overlapping, and that no single self-reported measure alone provides a better fit to HLA genetic ancestry than a combination including both race and geographic ancestry. We also found that patterns of reporting for race and ancestry appear to be influenced by participation in direct-to-consumer genetic ancestry testing.ConclusionsWhile these data are not used directly in matching for transplant, our results demonstrate that there is a place for the language of both race and geographic ancestry in the critical process of facilitating accurate prediction of HLA in the donor registry context.

Next-Generation Sequencing Reveals the Novel HLA-DQA1*06:01:04 Allele in a Bone Marrow Donor.

A single nucleotide mismatch within exon 2 of HLA-DQA1 differentiates the novel DQA1*06:01:04 allele from DQA1*06:01:01:01.

Identification of the Novel HLA-DQB1*03:01:01:69 Allele in a Candidate Bone Marrow Donor by Next Generation Sequencing.

The HLA-DQB1*03:01:01:69 allele differs from HLA-DQB1*03:01:01:03 by a single nucleotide substitution in intron 2.

Identification of the Novel HLA-DQB1*05:02:01:19 Allele in a Candidate Bone Marrow Donor by Next Generation Sequencing.

The HLA-DQB1*05:02:01:19 allele differs from HLA-DQB1*05:02:01:01 by a single nucleotide substitution in intron 3.

The Novel HLA-C*05:298 and HLA-DRB1*01:151 Alleles Identified in Russian Bone Marrow Donors.

Identification of two novel HLA alleles in Russian bone marrow donors.

Phase I/IIa Trial of Autologous Regulatory T Cell Therapy Together with Donor Bone Marrow Infusion in Kidney Transplantation

Phase I/IIa Trial of Autologous Regulatory T Cell Therapy Together with Donor Bone Marrow Infusion in Kidney Transplantation

Harnessing Hope in Digital Spaces for Health Equity: How Group Comparison Information and Supportive Comments Influence Bone Marrow Donor Intentions for African Americans.

Pursuing health equity necessitates recognizing health disparities that disproportionately impact disadvantaged groups and eliminating their barriers to essential health resources. Interactive digital technologies-specifically, popular social media platforms such as blogs and social networks-can be leveraged to engage underserved minority populations in collective social action aimed at addressing key determinants of health disparities and promoting equitable health outcomes. The present research focuses on the plight of African Americans-a minority group facing significant health disparities. Particularly in the domain of bone marrow donation, African Americans remain the group least likely to find a matching donor. Guided by the social comparison framing literature, we conducted an online experiment to investigate how group comparison information (GCI) emphasizing group-based disparities and supportive user comments on social media platforms influence African Americans' intentions to join a bone marrow registry. In doing so, we considered hope as a mediator and group identification as a moderator. Results based on a conditional process analysis showed that GCI led to greater bone marrow donor intentions in the presence of supportive comments through elicitation of hope, particularly among those low in group identification. The current findings demonstrate that it is important to consider the role of supportive message environments and group identification when addressing health disparities with GCI. Theoretical and practical implications are discussed.

Cryopreservation of hematopoietic cells from unrelated donors from the Czech National Bone Marrow Donor Registry in the context of COVID-19 pandemic – are there useless collections?

Cryopreservation of hematopoietic cells from unrelated donors from the Czech National Bone Marrow Donor Registry in the context of COVID-19 pandemic – are there useless collections?

HLA-A, -B, -C and -DRB1 Association with Autism Spectrum Disorder Risk: A Sex-Related Analysis in Italian ASD Children and Their Siblings.

Autism Spectrum disorders (ASD) are diagnosed more often in males than in females, by a ratio of about 3:1; this is likely to be due to a difference in risk burden between the sexes and/or to "compensatory skills" in females, that may delay the diagnosis of ASD. Identifying specific risk factors for ASD in females may be important in facilitating early diagnosis. We investigated whether HLA- class I: -A, -B, -C and class II -DRB1 alleles, which have been suggested to play a role in the development of ASD, can be considered as sex-related risk/protective markers towards the ASD. We performed HLA allele genotyping in 178 Italian children with ASD, 94 healthy siblings, and their parents. HLA allele distribution was compared between children with ASD, sex-matched healthy siblings, and a cohort of healthy controls (HC) enrolled in the Italian bone marrow donor registry. Allele transmission from parents to children with ASD and their siblings was also assessed. Our findings suggest that HLA-A*02, B*38, and C*12 alleles are more frequently carried by females with ASD compared to both HC and healthy female siblings, indicating these alleles as potential risk factors for ASD in females. Conversely, the HLA-A*03 allele was more commonly transmitted to healthy female siblings, suggesting it might have a protective effect. Additionally, the HLA-B*44 allele was found to be more prevalent in boys with ASD, indicating it is a potential risk factor for male patients. This is the first Italian study of sex-related HLA association with ASD. If confirmed, these results could facilitate early ASD diagnosis in female patients, allowing earlier interventions, which are crucial in the management of neurodevelopmental disorders.

Successful Allogeneic Hematopoietic Cell Transplantation for Patients with IL10RA Deficiency in Japan.

IL10RA (IL10 receptor subunit alpha) deficiency is an autosomal recessive disease that causes inflammatory bowel disease during early infancy. Its clinical course is often fatal and the only curative treatment is allogeneic hematopoietic cell transplantation (HCT). In Japan, only case reports are available, and there are no comprehensive reports of treatment outcomes. We retrospectively analyzed patients with IL10RA deficiency in Japan. Two newly identified and five previously reported patients were included in this study. Five patients underwent HCT; one untransplanted patient survived to age 14, and one died of influenza encephalopathy before transplantation. All five HCT recipients underwent HCT at the age before 2 years. They all were conditioned with fludarabine/busulfan- or fludarabine /melphalan-based regimens. The donor source was human leukocyte antigen haploidentical donor bone marrow (BM) for two patients and unrelated umbilical cord blood (CB) for two patients. One patient experienced graft failure with unrelated CB and required a second transplant with unrelated BM. All patients who underwent HCT survived and demonstrated an improved performance status. In cases of IL10RA deficiency, the need for transplantation should be promptly assessed, and early transplantation should be considered. (190/250).

A novel HLA-C*07 variant allele, HLA-C*07:01:01:141, identified by next-generation sequencing.

Characterisation of the novel HLA-C*07:01:01:141 allele in a 23-year-old Greek bone marrow donor.

Identification of the novel HLA-DQA1*03:79 allele in a candidate bone marrow donor by next generation sequencing.

A single nucleotide mismatch within exon 3 of HLA-DQA1 differentiates the novel allele DQA1*03:79 from DQA1*03:15.

Characterisation of the novel HLA-B*18:37:03 allele, first identified in a Brazilian individual.

The novel HLA-B*18:37:03 allele, first described in a potential bone marrow donor from Brazil.

361.9: Liver transplantation with delayed donor bone marrow transplantation in non-human primates achieves hematopoietic chimerism.

361.9: Liver transplantation with delayed donor bone marrow transplantation in non-human primates achieves hematopoietic chimerism.

Potential role of B- and NK-cells in the pathogenesis of pediatric aplastic anemia through deep phenotyping.

Pediatric patients with unexplained bone marrow failure (BMF) are often categorized as aplastic anemia (AA). Based on the accepted hypothesis of an auto-immune mechanism underlying AA, immune suppressive therapy (IST) might be effective. However, due to the lack of diagnostic tools to identify immune AA and prognostic markers to predict IST response together with the unequaled curative potential of hematopoietic stem cell transplantation (HSCT), most pediatric severe AA patients are momentarily treated by HSCT if available. Although several studies indicate oligoclonal T-cells with cytotoxic activities towards the hematopoietic stem cells, increasing evidence points towards defective inhibitory mechanisms failing to inhibit auto-reactive T-cells. We aimed to investigate the role of NK- and B-cells in seven pediatric AA patients through a comprehensive analysis of paired bone marrow and peripheral blood samples with spectral flow cytometry in comparison to healthy age-matched bone marrow donors. We observed a reduced absolute number of NK-cells in peripheral blood of AA patients with a skewed distribution towards CD56bright NK-cells in a subgroup of patients. The enriched CD56bright NK-cells had a lower expression of CD45RA and TIGIT and a higher expression of CD16, compared to healthy donors. Functional analysis revealed no differences in degranulation. However, IFN-γ production and perforin expression of NK-cells were reduced in the CD56bright-enriched patient group. The diminished NK-cell function in this subgroup might underly the auto-immunity. Importantly, NK-function of AA patients with reduced CD56bright NK-cells was comparable to healthy donors. Also, B-cell counts were lower in AA patients. Subset analysis revealed a trend towards reduction of transitional B-cells in both absolute and relative numbers compared to healthy controls. As these cells were previously hypothesized as regulatory cells in AA, decreased numbers might be involved in defective inhibition of auto-reactive T-cells. Interestingly, even in patients with normal distribution of precursor B-cells, the transitional compartment was reduced, indicating partial differentiation failure from immature to transitional B-cells or a selective loss. Our findings provide a base for future studies to unravel the role of transitional B-cells and CD56bright NK-cells in larger cohorts of pediatric AA patients as diagnostic markers for immune AA and targets for therapeutic interventions.

Chronic ethanol exposure decreases H3K27me3 in the Il6 promoter region of macrophages and generates persistent dysfunction on neutrophils during fungal infection.

The aim of this study was to investigate the effects of ethanol exposure on epigenetic markers in bone marrow (BM) and their impact on inflammatory response during Aspergillus fumigatus infection. Chronic ethanol exposure decreased H3K27me3 enrichment in the Il6 promoter region while increased H3K4me3 enrichment in Tnf. Chimeric mice were generated by transplanting BM from mice exposed to ethanol or water. Infection of ethanol-chimeric mice culminated in higher clinical scores, although there was no effect on mortality. However, previous chronic exposure to ethanol affects persistently the inflammatory response in lung tissue, demonstrated by increased lung damage, neutrophil accumulation and IL-6, TNF and CXCL2 production in ethanol-chimeric mice, resulting in a decreased neutrophil infiltration into the alveolar space. Neutrophil killing and phagocytosis were also significantly lower. Moreover, BM derived macrophages (BMDM) from ethanol-chimeric mice stimulated with A. fumigatus conidia exhibited higher levels of TNF, CXCL2 and IL-6 release and a higher killing activity. The Il6 promoter of BMDM from ethanol-chimeric mice exhibited a reduction in H3K27me3 enrichment, a finding also observed in BM donors exposed to ethanol. These evidences demonstrate that prior chronic alcohol exposure of bone-marrow modify immune effector cells functions impairing the inflammatory response during A. fumigatus infection. These findings highlight the persistent impact of chronic ethanol exposure on infectious disease outcomes.

In vitro evaluation of mesenchymal stromal cell senescence

Mesenchymal stromal cells (MSCs) are adult multipotent stem cells that, for their immunomodulatory and reparative/anti-inflammatory properties, are a promising tool in cell therapy. For the treatment of patients, MSCs are in vitro stimulated to proliferate in a non-physiological condition and this situation could lead MSCs to malignant transformation. The clinical application of MSCs requires that the biosafety of these cells must be investigated. The cellular senescence is a crucial mechanism that prevents the growth of cells at risk for neoplastic transformation. The aim of this study was to investigate the capacity of healthy donor bone marrow derived MSCs to enter the senescence phase by evaluating the β-galactosidase activity. We tested 131 MSC batches after long-term in vitro culture, resulting positive for β-gal staining and showing a branched shape morphology typical of senescent cells. They displayed a progressive decline in proliferative capacity and none of them bypassed the senescence. In conclusions, our data indicate that MSCs are not inclined towards spontaneous neoplastic transformation. We believe that the control of MSC capacity to enter senescence is fundamental for quality and safety, considering the relevant interest in the MSC clinical applications.