Cord Blood Stem Cells Research Articles

Evaluation the Impressionability of Acellular Scaffolds in Presence of Different Combination of Umbilical Cord Blood Stem Cells and Platelet Rich Fibrin in Repair of Knee Defects in Rabbit (Novel Method with Xeno-Material Elements

Background: Currently, the use of xenogeneic and allogeneic compounds in the treatment of musculoskeletal diseases is common. Repairing bone and cartilage remains a significant challenge for orthopedic surgeons. One of the principal goals of tissue engineering is the development of appropriate scaffolds that can promote tissue regeneration. Various cells and genes are involved in bone and cartilage regeneration, and new scaffolds can induce these processes during osteoregeneration. Objectives: The aim of this study is to evaluate the effects of umbilical cord blood stem cells (USCs) and platelet-rich fibrin (PRF) using acellular scaffolds in the repair of knee defects in rabbits. Methods: In this study, the femoral patella of 12 male New Zealand rabbits was drilled, and they were divided into four groups: (1) control (rabbits with femoral defects), (2) acellular osteochondral (AO), (3) AO + PRF, and (4) AO + USCs / platelet-rich fibrin. Different scaffolds were implanted into their knees, and after six months, histological evaluations were conducted. To further investigate the effects of scaffolds on bone and cartilage, gene expression levels of Col 1, Col X, Runx2, SOX9, and ALP were measured using real-time PCR. Results: All the implanted materials contributed to knee repair. In terms of statistical analysis, the use of USCs and platelet-rich fibrin with natural scaffolds such as Acellular Osteochondral provided better results for repairing bone and cartilage. The evaluation of specific bone regeneration genes (COLI, RUNX2) and the histological results from the implanted site in experimental osteochondral defects indicated that the most effective knee repair occurred in the group treated with cell-free osteochondral scaffolds, USCs, and platelet-rich fibrin. Conclusions: This study demonstrates that the combination of biomaterials and xenografts can accelerate the regeneration process.

A DESCRIPTIVE STUDY TO ASSESS KNOWLEDGE REGARDING UMBILICAL CORD BLOOD BANKING AMONG WOMEN IN THE REPRODUCTIVE AGE

Umbilical cord blood stem cells are the name given to the blood remaining in the umbilical cord and placenta after childbirth. Umbilical cord blood has the potential to be a huge source of primitive hematopoietic stem and progenitor cells that can be used to reconstruct the hematopoietic system and restore immunological function in individuals who require treatment. A descriptive research methodology was adopted in this study and 100 reproductive age group women in Sharda Hospital were selected in the present study. Convenience sampling technique was adopted for the data collection. The information was gathered via a knowledge questionnaire, which was used to assess knowledge regarding umbilical cord blood banking. The result revealed that 30 women had poor knowledge, 63 women had moderate knowledge and 7 had knowledge about umbilical cord blood banking. The importance of umbilical cord blood banking information was given among selected 100 participating women via the help of a pamphlet to raise awareness of uses of umbilical cord bloodbanking.

TO ASSESS THE EFFECTIVENESS OF VIDEO ASSISTED TEACHING PROGRAMME ON KNOWLEDGE REGARDING IMPORTANCE OF UMBILICAL CORD BLOOD PRESERVATION FOR FUTURE TREATMENT MODALITIES AMONG EXPECTANT COUPLES AT SELECTED HOSPITALS

Health is the general condition of a person in all aspects it is also a level of functional and metabolic efficiency of an organism. Nothing is more important than protecting the health of your family. A healthy future starts with saving - or banking - your newborns cord blood stem cells. Its a one-time opportunity that can change or even save a life. After a baby is delivered, the mothers body releases the placenta, the temporary organ that transferred oxygen and nutrients to the baby while in the mothers uterus. Stem cell research holds enormous promise for easing human suffering. We have a lot to gain through furthering stem cell research, but medical breakthroughs should be fundamentally about saving, not destroying, human life. Therefore, support stem cell research that does not destroy the embryo. The aim of this study is to assess the effectiveness of video assisted teaching programme on knowledge regarding importance of umbilical cord blood preservation for future treatment modalities among expectant couples. The main objective of the study was to assess the knowledge regarding importance of umbilical cord blood preservation for future treatment modalities among expectant couples. Methodology: The sample size was 60 expectant couples and the setting of the study was selected hospitals where the couples come for antenatal checkup. Sampling technique was convenient sampling technique. One group pretest posttest design was used. Result: The study shows that in pre-test 81.67% of couples were having inadequate knowledge and 18.33% of them having moderately adequate knowledge and none of them having adequate knowledge. While in post test 55% of expectant couples were having Moderately Adequate knowledge and 45% of them having adequate knowledge and none of them having inadequate knowledge. The result shows that the video assisted teaching was effective in improving the knowledge. There was significant association between selected demographic variables and post test score. Conclusion: The teaching was effective in increasing the knowledge of expectant couples and similar awareness programmes should be given to the public so that they can understand regarding the newer treatment modalities.

Gestational diabetes mellitus affects the differentiation of hematopoietic stem cells in neonatal umbilical cord blood.

There are abundant hematopoietic stem cells (HSCs) in cord blood. It is known that HSCs continue to differentiate to CLP, CMP and erythroid progenitor cells (EPC), EPC ultimately differentiated to platelets and erythrocytes. It has been reported that the proportion of HSCs in cord blood was higher than that in healthy pregnant women, so as the incidence of neonatal polycythemia in gestational diabetes mellitus (GDM) patients. We aimed to investigate whether the hyperglycemic and/or hyperinsulin environment in GDM patients has effects on the differentiation of HSCs into erythrocytes in offspring cord blood. In this study, we collected cord blood from 23 GDM patients and 52 healthy pregnant women at delivery. HSCs, CLP, CMP and EPCs in cord blood of the two groups were identified and quantified by flow cytometry. HSCs were sorted out and treated with glucose and insulin, respectively, and then, the changes of HSCs proliferation and differentiation were detected. Compared to healthy controls, HSCs, CMP and EPC numbers in cord blood from GDM group were significantly increased, while CLP cell number was decreased. The differentiation of HSCs into EPC was promoted after treatment with glucose or insulin. There were more HSCs in the cord blood of GDM group, and the differentiation of HSCs to EPCs was increased. These findings were probably caused by the high-glucose microenvironment and insulin medication in GDM patients, and the HSCs differentiation changes might be influencing factors of the high incidence of neonatal erythrocytosis in GDM patients.

Umbilical cord blood stem cells: transplantation prospects in neurological practice

Umbilical cord blood has evolved from being considered a mere cellular/tissue waste product after childbirth to a valuable biological material with a wide regenerative potential. Today, hematopoietic cord blood stem cells are used in the same way as bone marrow and mobilised peripheral blood as part of the standard medical treatment for haemoblastoses and hereditary blood diseases. There is an increasing amount of experimental data showing possibility of umbilical cord blood cell fractions using for treatment of non-hematological diseases and, in particular, diseases of the central nervous system. One of the key challenges in cell therapy for central nervous system diseases is the choice of cellular material for neurotransplantation. This is particularly relevant for ischemic and traumatic injuries, as well as neurodegenerative diseases. Stem or mature somatic cells prepared for neurotransplantation should have predictable and reproducible characteristics thataccording therapeutic purposes, exactly: trophic and/or neuroprotective action to increase neurons viability in damaged area; axon growth stimulation and myelination; cell matrix restoration or predictable differentiation direction to replace amount of lost brain or spinal cord cells. The fact that umbilical cord blood contains not only hematopoietic, but also various non-hematopoietic stem cells with a wide regenerative potential has become the basis for active use of these cells for neurotransplantation. The proposed review provides historical information about umbilical cord blood introduction into practical medicine, its cellular composition and clinical applications of stem cells various types for children central nervous system diseases treatment, such as ischemic encephalopathy, stroke, cerebral hemorrhage, cerebral palsy, autism, and in adults — for stroke treatment, spinal cord injury and neurodegenerative diseases.

Characterization of cord blood immune and stem cells from Vietnamese infants

Cord blood has emerged as a valuable and versatile resource, holding within its grasp a myriad of immune and stem cell types with immense therapeutic potential. Moreover, immune profiling in cord blood, a burgeoning field of research, offers profound insight into the early immune development of newborns, paving the way for understanding immune health, disease susceptibility, and therapeutic interventions. Therefore, in this research, we aimed to characterize the diverse immune and stem cell types present in the cord blood of Vietnamese infants. In this study, a total of 59 cord blood samples were examined for immune cell populations. Cell populations were identified using flow cytometry based on the expression of specific markers on the cell surface. The results showed that the CD3+/CD4+ population accounted for the greatest proportion of lymphocytes (46.71 ± 8.99%), followed by the CD3+/CD8+ population (22.58 ± 6.73%). The mature B-cell population (CD3-/CD19+) accounted for 9.52 ± 3.28%, while CD3+/CD19+ cells accounted for an average of 8.59 ± 5.77%. Moreover, the NK population (CD3-/CD56+ cells) accounted for 6.44 ± 4.9%, T lymphocytes carrying both CD4 and CD8 markers (CD3+/CD4+/CD8+) accounted for 0.63 ± 0.41%, and NKT cells (CD3+/CD56+) accounted for only 0.30 ± 0.53%. Moreover, the number of hematopoietic stem cells (HSCs) tends to increase in umbilical cord blood units with a high volume and number of TNCs and MNCs. In contrast, the number of HSCs was negatively correlated with the ratio of B lymphocytes. The TCD4 ratio was positively correlated with the number of HSCs but negatively correlated with the NK ratio (p = 0.01). The study initially characterized the diversity of some stem and immune cell types present in the umbilical cord blood of Vietnamese infants. These findings laid the groundwork for developing novel immune profiles and expanding the potential applications of umbilical cord blood in cellular therapy.

Knowledge and attitude on umbilical cord blood stem cell therapy among bachelor nursing students in Lalitpur Metropolitan City

Background: Stem cells are immature cells that have ability to mature into different types of functioning cells as needed by body. Umbilical cord blood is one of the richest sources of stem cells. It is vital for nurses to keep up-to-date of current advances in stem cell science and nurses can play crucial role in providing first-hand information to public about stem cell and umbilical cord blood banking. Therefore, this study is aimed to identify the knowledge and attitude regarding umbilical cord blood stem cell therapy among bachelor nursing students. Method: A descriptive cross sectional research design was used to assess the knowledge and attitude of 263 nurses studying bachelor in nursing science and post basic bachelor in nursing in different selected nursing college of Lalitpur metropolitan city. Multi stage cluster sampling was used to select respondents. Data was collected using self-developed self-administered google form questionnaire. Results: Most of the nurses had poor knowledge (49.4%) and few nurses had good knowledge (6.8%) however majority of nurses (93.9%) had favourable attitude regarding umbilical cord blood stem cell therapy. Regarding association between variable, there is no any association between knowledge level and attitude (p=0.224). Conclusion: Nurses had poor knowledge and favourable attitude regarding umbilical cord blood stem cell therapy. Keywords: Nurses,Stem cells, Umbilical cord blood

Desensitization therapy for elevated donor-specific antibody levels in haploidentical transplantation in Chilean patients

Allogeneic stem cell transplantation (ALOSCT) is curative for several hematological diseases. Unrelated donors and cord blood stem cells are valid options but haploidentical donors (HAPLO) have been considered the main source of stem cells in several countries, partly due to easy access and low cost of the transplantation process. Unfortunately, some patients have antibodies against the HLA epitopes from family haploidentical donors (donor-specific antibodies [DSA]) which are associated with engraftment failure and lethality. A few strategies exist to reduce or eliminate HLA antibodies that bind to these receptors. In this study, we present our experience with DSA desensitization by retrospectively examining a cohort from our hospital program. Between 2012 and 2023, we performed 243 ALOSCTs, of which 142 were from HAPLO and 56 were from unrelated donors. Nine patients (7%) had elevated DSA levels, most of which were female patients and mostly HAPLO. The median fluorescence intensity for these patients was 22,490 (19,000-28,560). Most of these high DSA patients (80%) received a desensitization procedure that involves plasmapheresis, rituximab, and immunoglobulin. The remaining 20% had severe infections during transplantation, and received rituximab monotherapy instead. The median dose of stem cells infused was 6.5 x 10^6 CD34/kg. Graft-versus-host disease (GVHD) preventative measures for all patients involved post-transplantation cyclophosphamide. Primary graft failure was observed in 45% of DSA elevated patients. For the remaining patients, median granulocyte and platelet engraftment were 14 d (12-16) and 16 d (13-18), respectively. Mortality in patients who did not receive engraftment was 100%. The incidence of mild, chronic GVHD was 15%. In conclusion, the desensitization of DSA in patients provided a 55% rate of engraftment and survival. However, a 45% rate of primary graft failure continued to pose a challenge in patients with DSA and required the development of improved strategies to reduce elevated transplant-related mortality.

Role of Human Umbilical Cord Blood-derived Stem Cells versus their Conditioned Medium on the Regeneration of Pancreatic Beta Cells in a Rat Model of Diabetes Mellitus

Role of Human Umbilical Cord Blood-derived Stem Cells versus their Conditioned Medium on the Regeneration of Pancreatic Beta Cells in a Rat Model of Diabetes Mellitus

Effect of fetal distress on viability and yield of umbilical cord blood stem cells–a prospective comparative study

IntroductionDifferent factors affect the quality and viability of cord blood stem cells, and therefore the efficacy of umbilical cord stem cell transplantation. Fetal distress is one factor affecting the quantity of CD34+ cells in cord blood. This study was designed to compare the viability and yield of the umbilical cord blood stem cells of women who have undergone emergency lower segment caesarean section for fetal distress or for other causes. Materials and methodsThis cross-sectional analytical study was performed at a tertiary care hospital facility with a total sample size of 68: 34 participants had undergone emergency C-section for fetal distress, and 34 had undergone emergency C-section for other causes. Umbilical cord blood was collected ex-utero in a 350 mL bag with citrate-phosphate-dextrose solution with adenine. Three milliliter of blood were transferred to an ethylenediaminetetraacetic acid (EDTA) tube for cell counts and flow cytometry testing for CD34+. The chi-square test was used to compare the total mononuclear cell, CD34+, and viability between the groups. ResultsThe CD34+ count [mean 4.9 versus 1.1 (× 106 cells/unit)] and total nucleated cell count [mean 14.2 versus 7.5 (× 108/unit)] were significantly higher in cord blood units collected from women who delivered by C-section for fetal distress (p-value <0.05). However, the volume of umbilical cord stem cells and viability of stem cells did not vary significantly based on the presence or absence of fetal distress (p-value >0.05). ConclusionThe current study shows that umbilical cord blood collected during fetal distress has a significantly higher content of stem cells and total nucleated cells than the non-fetal distress group

The Evaluation of Mass/DNA Copy Number of Mitochondria in Umbilical Cord Blood-derived Hematopoietic Stem Cells Cocultured with MSCs.

Over recent decades, UCB has been widely used as an excellent alternative source of HSCs for treating many hematologic disorders. Recent studies suggest using mesenchymal stroma cell co-cultures to increase the number of HSCs prior to transplantation. Considering the critical role of mitochondria in the cell's fate and the importance of the self-renewal capacity of HSCs in HSCT, we decided to investigate the mass/DNA copy number of mitochondria in HSCs while co-cultured with MSCs and alone after seven days. UCB units were collected from full-term deliveries. MSCs and HSCs were isolated from UCB and the purity of cells was confirmed by flow cytometry. The mtDNA-Copy Number of HSCs was calculated using prob-based real-time PCR. Furthermore, Mito Tracker Green dye measured the mass of mitochondria of HSCs. HSCs from MSC co-culture group showed significantly fewer mtDNA-CN compared to HSCs alone after seven days (p < 0.001). Besides, by comparing the two groups on day seven to HSCs on day zero, we observed a mild increase in the mitochondrial mass of HSCs alone compared to the MSC-HSC co-culture group (p < 0.05). Concerning previous studies that have proved the association between lower mass/DNA-copy number of mitochondria in CD34 + HSCs and lower metabolic activity along with higher quiescence maintenance, and by considering the results of this experiment, it seems that the MSC-HSC co-cultures might be associated with a higher expansion of HSCs as well as stemness maintenance leading to the improvement in engraftment. Nevertheless, further investigations are required to clarify the exact connection between lower mass/DNA-copy number of mitochondria and stemness maintenance in HSCs.

The Ethical Challenges of Umbilical Cord Blood Stem Cell Banking: A Qualitative Study

Background: Umbilical cord blood serves as a potent source of hematopoietic stem cells, utilized in treating specific diseases and preserved in blood banks. Objectives: This study aims to explore the ethical issues associated with these repositories. Methods: This qualitative research involved conducting 14 interviews with experts and the families of recipients or donors of umbilical cord blood. Participants were selected purposefully from May to November 2021. Results: The investigation identified five principal ethical challenges: presentation of information, interpersonal relationships among involved parties, respect for human dignity, adherence to rules and regulations, and the handling of tissue samples. These challenges encompassed various categories and subcategories, including informed consent, communication and advertising of information, financial considerations, conflicts of interest, professional interactions, ownership rights concerning the child and parents, privacy and confidentiality issues, compliance with regulations and guidelines, and the clinical and research uses of the tissue samples. Conclusions: The study unveiled significant ethical concerns in the domain of umbilical cord blood stem cell banking. Addressing these ethical dilemmas necessitates the involvement of health policymakers and medical ethics experts, along with a comprehensive understanding of these banks' multifaceted nature by the community.

Abstract 2783: The exosomes from cord blood stem cells, containing novel miRNAs, induce apoptosis in melanoma cells and enhance T cell anticancer function

Abstract Regulatory T (Treg) cells expressing FOXP3 play a critical role in suppressing immune responses against anti-tumor immune responses. However, their presence in tumor tissues is often associated with poor prognosis. Cord blood stem cell-derived exosome vesicles (CBSC-EV) constitute a valuable source of lipids, microRNAs, and proteins, that are essential for cell-to-cell communication and provide therapeutic benefits. This study aimed to investigate the contents of CBSC-EV, identifying several known miRNAs (from Let7 and Let5 families) and two unknown miRNAs. Healthy lymphocytes, fibroblasts, and CHL1 amelanotic melanoma cell lines were transfected with these two novel miRNA inhibitors, and the changes before and after treatment compared to CBSC-EV and H2O2 were examined. Following treatment, the cells underwent a 24-hour incubation period, during which the level of cytotoxicity was assessed using CCK8, and the degree of DNA damage incurred was quantified with the Fast Microplate DNA damage assay. To further investigate the effects of treatment, RNA sequencing was conducted on both healthy and cancerous cells. The results demonstrated that CBSC-EV and novel miRNAs induced cytotoxicity in cancer cells, while in healthy cells, the DNA damage was repaired and cell viability was significantly increased (p&amp;lt;0.001). RNA sequencing revealed that following treatment with CBSC-EV and both CBSC-EV+N1 and N2 inhibitors in malignant melanoma cells, the expression of genes involved in the cytokine-mediated signaling pathway, cytokine-cytokine interaction, and cytokine activity were enhanced. In healthy cells treated with CBSC-EV and novel miRNA inhibitor transfection, the MT-CO1 gene was upregulated, contributing to cytochrome-c oxidase activity. This study presents evidence of the potential use of novel miRNAs as genetic material for cancer therapeutics in the future. Additionally, the findings suggest that these miRNAs could be used as an immune system modulator against melanoma. These findings demonstrate that miRNAs can effectively suppress resistance to anticancer cytotoxic therapy, which is a common feature of cancer cells. This suggests that miRNAs can potentially enhance current cancer therapies. Citation Format: Mojgan Mojgan Najafzadeh, Adi Baumgartner, Shohreh Jafarinejad, Zahra Karimi, Mohammad Isreb, Pouria Akhbari, Nader Ghaderi, Farshid Sefat, Saeed Heidari Keshel, Parisa Naeem, Rojan Ghaderi, Jacobo Elis Gomez, Diana Anderson, Andrew Wright. The exosomes from cord blood stem cells, containing novel miRNAs, induce apoptosis in melanoma cells and enhance T cell anticancer function [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2783.

Abstract 1420: Mechanism of NPRL2 gene therapy induced antitumor immunity in KRAS/STK11mt aPD1 resistant metastatic NSCLC

Abstract NPRL2/TUSC4 is a tumor suppressor gene whose expression is reduced in many cancers including NSCLC. Restoration of NPRL2 induces cell cycle arrest and apoptosis. We investigated the antitumor immune responses to NPRL2 gene therapy in aPD1 resistant KRAS/STK11mt NSCLC in a humanized mouse model. Humanized mice were generated by transplanting human cord blood-derived CD34 stem cells into NSG mice. Mice harboring &amp;gt; 25% human CD45 cells were considered humanized. Lung metastases were developed in humanized mice by injecting KRAS/STK11mt/aPD1R A549 cells, which were treated with i.v. injection of NPRL2 nanoparticles (DOTAP-NPRL2) with or without pembrolizumab (aPD1). NPRL2 treatment reduced lung metastases, whereas pembrolizumab was ineffective. The antitumor effect was greater in humanized than non-humanized mice suggesting the role of immune cells. The antitumor effect was associated with increased infiltration of human cytotoxic immune cells and decreased numbers of Treg in tumors. NPRL2+ pembrolizumab was not synergistic in this resistant model but was synergistic in KRASwt/aPD1S H1299 tumors. Cytotoxic immune cells in tumors were associated with the antitumor effect. Consistent with A549, NPRL2 showed a significantly strong antitumor effect on other KRASmt/aPD1R syngeneic LLC2 tumors whereas aPD1 was not effective. The antitumor effect of NPRL2 was correlated with HLA-DR+ DC, CD11c DC, TILs and NK cells in TME. The antitumor effect of NPRL2 was abolished upon in-vivo depletion of CD4, CD8 T, and MΦ in the LLC2 tumor model. However, no effect was found on in-vivo depletion of NK cells. Nanostring analysis on lung metastasis resulted in a distinct pattern of human gene expression by NPRL2. T cell functional genes, including IFNγ, CD8b, CD7, TNFSF18, ITGA1, GATA3 and TBX21 were significantly increased. Conversely, the negative regulatory genes, including FOXP3, TGFB1, TGFB2, and IL-10RA were inhibited. NPRL2 also downregulated T cell co-inhibitory molecules, including CTLA4, ICOS, LAG3, PDCD1, CD274, IDO1, PDCD1LG2, CD47, and KLRB1. Stably expressing NPRL2 clones were established, and tumors in humanized mice with these clones exhibited significantly slower growth compared to controls. TME analysis showed an upregulation of human CD45, CD3, CD8 T, HLA-DR+ DC and a downregulation of Tregs, CD3+PD1+T, MDSC, and CD163+ TAM in tumors expressing NPRL2. The stable cells showed a substantial increase in both colony formation inhibition and apoptosis. Stable clones showed heightened sensitivity to carboplatin in colony formation, apoptosis, and PARP cleavage assays. Stable expression of NPRL2 resulted in the downregulation of both AKT-mTOR and MAPK pathways by inhibition of pAKT, pmTOR, pPRAS40, p4E-BP1, pS6, and pERK1/2. Taken together, NPRL2 gene therapy induces antitumor activity on KRAS/STK11mt/aPD1R tumors through DC-mediated antigen presentation and cytotoxic immune cell activation Citation Format: Ismail M. Meraz, Mourad Majidi, Renduo Song, Feng Meng, Lihui Gao, Qi Wang, Jing Wang, Elizabeth Shpall, Jack A. Roth. Mechanism of NPRL2 gene therapy induced antitumor immunity in KRAS/STK11mt aPD1 resistant metastatic NSCLC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1420.

Abstract 4493: Protein structure inspired drug discovery

Abstract Introduction: Drug action is mediated by a small molecule therapeutic binding to a specific structural motif on a protein. As structure determines function, it follows that unique structural motifs are enriched for sites of unique function and high therapeutic target value. We hypothesized that protein structure constitutes high value primary information that can drive the discovery of novel therapeutic agents. Method: We developed a unique physics-based protein structure analysis platform scaled to run on a supercomputer, allowing us to consider dynamic protein flexibility and the influence of solvent upon protein structure, and to probe large protein libraries as a primary source of structure information that can support up-front structure-based screens. We developed and then probed a high resolution protein x-ray crystallographic library, created and curated by us. From a 60 million compound library, we conducted virtual screens for docking to identified sites. 8-day colony formation assays were conducted on a panel of 8 human breast, prostate, lung and colon cancer cell lines, as well as 8- and 14-day trilineage hematopoietic colony formation assays on human cord blood stem cells. Findings: From the first 180 deposited protein structures, we screened for unique protein surface structure. Structures were deemed unique if they were not otherwise known to be associated with any particular functional roles and were not the binding site of known therapeutics. Further selection included those that were clefts and possessed physical characteristics compatible with binding small chemicals whose properties have been linked to effective therapeutics. For each of 8 sites on 6 different proteins, a suite of analytics probed docking solutions from the compound library, generating a ranked list of 100 compounds for each site. Based on factors inclusive of availability and low potential for toxicity, compounds were acquired and used in in-parallel selection and de-selection functional assays. Cancer cell growth inhibition was a positive selection criterion, with additional ranking based on greater potency and broader efficacy. Bone marrow toxicity is limiting for most drugs, and inhibition of bone marrow stem cell growth was as a deselection criterion. Multiple compounds clustered around 2 sites on 2 different proteins. One compound, Dxr2-017, exhibited very high relative efficacy against colon and breast cancer cells. Evaluation of Dxr2-017 in the NCI-60 panel corroborated these findings, and showed even higher efficacy against melanoma. We demonstrated that Dxr2-017 inhibited melanoma growth with low nanomolar efficacy, while concentrations of over 2300-fold higher had only limited inhibitory effects on bone marrow stem cells. Conclusions: This study provides proof-of-principle that protein structure constitutes high value primary information that can support identification of novel therapeutics. This approach is widely applicable and expandable. Citation Format: Fangfang Qiao, T. Andrew Binkowski, Wayne Anderson, Weining Chen, Gray Schiltz, Karl Scheidt, Amarnath Natarajan, Raymond Bergan. Protein structure inspired drug discovery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4493.

Therapeutic effect and study of human umbilical cord blood mononuclear cells in patients with ischaemic bowel disease

Ischaemic bowel disease (ICBD) is a group of intestinal ischaemia syndromes caused by various aetiologies of reduced intestinal blood flow or vascular occlusion. ICBD can present as abdominal pain, bloody stool, and diarrhoea. This disease often occurs in middle-aged and elderly individuals with cardiovascular and cerebrovascular diseases. The incidence of ischaemic bowel disease has been increasing for decades, and it is difficult to diagnose, resulting in rapid disease progression and a high mortality rate. Therefore, fully understanding this disease, improving the diagnosis rate of this disease, and finding appropriate treatment methods are urgently needed to improve the condition and prognosis of patients. Umbilical cord blood stem cells are accessible, have weak immunogenicity, and have various biological functions, such as angiogenesis, inflammation and immune regulation. Many studies have confirmed that cord blood stem cells can relieve ischaemia, and these cells have attracted tremendous amounts of attention in regenerative medicine in recent years. In this paper, we discuss the clinical characteristics of ICBD, analyse the characteristics of human umbilical cord blood mononuclear cells (HUCB-MNCs), and use its to treat ischaemic bowel disease. Additionally, we compare the clinical manifestations and related indicators before and after treatment to evaluate the efficacy and safety of these methods.

Effects of different concentrations of nicotinamide on hematopoietic stem cells cultured in vitro.

In vitro expansion to increase numbers of hematopoietic stem cells (HSCs) in cord blood could improve clinical efficacy of this vital resource. Nicotinamide (NAM) can promote HSC expansion ex vivo, but its effect on hematopoietic stem and progenitor cells (HSPCs, CD34+CD38) and functional subtypes of HSCs - short-term repopulating HSCs (ST-HSCs, CD34+CD38CD45RACD49f+) and long-term repopulating HSCs (LT-HSCs, CD34+CD38CD45RACD49f+CD90+) is not yet known. As a sirtuin 1 (SIRT1) inhibitor, NAM participates in regulating cell adhesion, polarity, migration, proliferation, and differentiation. However, SIRT1 exhibits dual effects by promoting or inhibiting differentiation in different tissues or cells. We propose that the concentration of NAM may influence proliferation, differentiation, and SIRT1 signaling of HSCs. To evaluate the effects and underlying mechanisms of action of different concentrations of NAM on HSC proliferation and differentiation. CD34+ cells were purified from umbilical cord blood using MacsCD34 beads, and cultured for 10-12 d in a serum-free medium supplemented with cytokines, with different concentrations of NAM added according to experimental requirements. Flow cytometry was used to detect phenotype, cell cycle distribution, and apoptosis of the cultured cells. Real-time polymerase chain reaction was used to detect the transcription levels of target genes encoding stemness-related factors, chemokines, components of hypoxia pathways, and antioxidant enzymes. Dichloro-dihydro-fluorescein diacetate probes were used to evaluate intracellular production of reactive oxygen species (ROS). Determination of the effect of different culture conditions on the balance of cytokine by cytometric bead array. Compared with the control group, the proportion and expansion folds of HSPCs (CD34+CD38) incubated with 5 mmol/L or 10 mmol/L NAM were significantly increased (all P < 0.05). The ST-HSCs ratio and fold expansion of the 5 mmol/L NAM group were significantly higher than those of the control and 10 mmol/L NAM groups (all P < 0.001), whereas the LT-HSCs ratio and fold expansion of the 10 mmol/L NAM group were significantly higher than those of the other two groups (all P < 0.05). When the NAM concentration was > 10 mmol/L, cell viability significantly decreased. In addition, compared with the 5 mmol/L NAM group, the proportion of apoptotic cells in the 10 mmol/L NAM group increased and the proportion of cells in S and G2 phase decreased. Compared with the 5 mmol/L NAM group, the HSCs incubated with 10 mmol/L NAM exhibited significantly inhibited SIRT1 expression, increased intracellular ROS content, and downregulated expression of genes encoding antioxidant enzymes (superoxide dismutase 1, peroxiredoxin 1). Low concentrations (5 mmol/L) of NAM can better regulate the balance between proliferation and differentiation, thereby promoting expansion of HSCs. These findings allow adjustment of NAM concentrations according to expansion needs.

Haploidentical donor peripheral blood stem cell transplantation using third-party cord blood compared with matched unrelated donor transplantation for patients with hematologic malignancies

Objectives: To assess the efficacy of cord blood-assisted haploid peripheral blood stem cell transplantation (haplo-cord-PBSCT) versus unrelated donor peripheral blood stem cell transplantation (UD-PBSCT) in the treatment of malignant hematological diseases. Methods: A retrospective analysis was performed on one hundred and four patients with malignant hematological diseases who underwent haplo-cord-PBSCT and fifty-two patients who underwent UD-PBSCT at Xiangya Hospital of Central South University between January 2016 and December 2021. Results: ①The median implantation time for neutrophils in the haplo-cord-PBSCT and UD-PBSCT groups was 13 (9-22) days and 13 (10-24) days, respectively (P=0.834), whereas the median implantation time for platelets was 15 (7-103) days and 14 (8-38) days, respectively (P=0.816). The cumulative implantation rate of neutrophils at 30 days after transplantation in the haplo-cord-PBSCT group and the UD-PBSCT group was 100% (P=0.314), and the cumulative platelet implantation rate at 100 days after transplantation was 95.2% (95% CI 88.3% - 98.1% ) and 100% (P=0.927), respectively. 30 days after transplantation, both groups of patients achieved complete donor chimerism, and no umbilical cord blood stem cells were implanted. ②The cumulative incidence rates of grade Ⅱ-Ⅳ acute GVHD within 100 days after transplantation in the haplo-cord-PBSCT group and the UD-PBSCT group were 29.1% (95% CI 20.1% -38.1% ) and 28.8% (95% CI 17.2% -41.6% (P=0.965), respectively. The cumulative incidence rates of grade Ⅲ/Ⅳ acute GVHD were 7.8% (95% CI 3.6% -14.0% ) and 9.6% (95% CI 3.5% -19.5% ) (P=0.725). The cumulative incidence rates of 2-year chronic GVHD in the haplo-cord-PBSCT group and the UD-PBSCT group were 45.3% (95% CI 36.1% -56.1% ) and 35.1% (95% CI 21.6% -44.1% ), respectively (P=0.237). The cumulative incidence rates of severe chronic GVHD at 2 years after transplantation were 13.6% (95% CI 7.6% -21.3% ) and 12.9% (95% CI 5.1% -24.3% ), respectively (P=0.840). ③The 2-year CIR after transplantation in the haplo-cord-PBSCT group and UD-PBSCT group were 12.8% (95% CI 7.0% -20.5% ) and 10.0% (95% CI 3.6% -20.2% ), respectively (P=0.341), and the NRM were 14.7% (95% CI 8.4% -22.6% ) and 16.2% (95% CI 7.4% -28.0% ), respectively (P=0.681). ④The 2-year OS rates in the haplo-cord-PBSCT and UD-PBSCT groups after transplantation were 82.2% (95% CI 74.8% -90.3% ) and 75.5% (95% CI 64.2% -88.7% ), respectively (P=0.276). The 2-year DFS rates were 69.9% (95% CI 61.2% -79.8% ) and 73.8% (95% CI 62.4% -87.3% ), respectively (P=0.551). The 2-year rates of GVHD-free/recurrence-free survival (GRFS) were 55.3% (95% CI 44.8% -64.8% ) and 64.7% (95% CI 52.8% -79.3% ), respectively (P=0.284) . Conclusion: The findings of this study indicate that haplo-cord-PBSCT and UD-PBSCT have comparable efficacy and safety in the treatment of malignant hematological diseases and can be used as an alternative treatment options.

Suppression of B-Cell Activation by Human Cord Blood-Derived Stem Cells (CB-SCs) through the Galectin-9-Dependent Mechanism.

We developed the Stem Cell Educator therapy among multiple clinical trials based on the immune modulations of multipotent cord blood-derived stem cells (CB-SCs) on different compartments of immune cells, such as T cells and monocytes/macrophages, in type 1 diabetes and other autoimmune diseases. However, the effects of CB-SCs on the B cells remained unclear. To better understand the molecular mechanisms underlying the immune education of CB-SCs, we explored the modulations of CB-SCs on human B cells. CB-SCs were isolated from human cord blood units and confirmed by flow cytometry with different markers for their purity. B cells were purified by using anti-CD19 immunomagnetic beads from human peripheral blood mononuclear cells (PBMCs). Next, the activated B cells were treated in the presence or absence of coculture with CB-SCs for 7 days before undergoing flow cytometry analysis of phenotypic changes with different markers. Reverse transcription-polymerase chain reaction (RT-PCR) was utilized to evaluate the levels of galectin expressions on CB-SCs with or without treatment of activated B cells in order to find the key galectin that was contributing to the B-cell modulation. Flow cytometry demonstrated that the proliferation of activated B cells was markedly suppressed in the presence of CB-SCs, leading to the downregulation of immunoglobulin production from the activated B cells. Phenotypic analysis revealed that treatment with CB-SCs increased the percentage of IgD+CD27- naïve B cells, but decreased the percentage of IgD-CD27+ switched B cells. The transwell assay showed that the immune suppression of CB-SCs on B cells was dependent on the galectin-9 molecule, as confirmed by the blocking experiment with the anti-galectin-9 monoclonal antibody. Mechanistic studies demonstrated that both calcium levels of cytoplasm and mitochondria were downregulated after the treatment with CB-SCs, causing the decline in mitochondrial membrane potential in the activated B cells. Western blot exhibited that the levels of phosphorylated Akt and Erk1/2 signaling proteins in the activated B cells were also markedly reduced in the presence of CB-SCs. CB-SCs displayed multiple immune modulations on B cells through the galectin-9-mediated mechanism and calcium flux/Akt/Erk1/2 signaling pathways. The data advance our current understanding of the molecular mechanisms underlying the Stem Cell Educator therapy to treat autoimmune diseases in clinics.

Effect of allogeneic hematopoietic stem cell transplantation for chronic granulomatous disease in children: A multicentre, retrospective cohort study in China

Chronic granulomatous disease (CGD) in children is a rare primary immunodeficiency disorder that can lead to life-threatening infections and inflammatory complications. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is increasingly being used to treat severe CGD in children. We conducted a multicenter retrospective analysis of children with CGD who were treated with allo-HSCT at four pediatric hematopoietic stem cell transplant centers in China from September 2005 to December 2019. The study included a total of 171 patients (169 males and 2 females). The median age at the time of transplantation was 6.1 (0–16.4) years. Among them, 154 patients had X-linked recessive inheritance caused by CYBB gene mutations, 12 patients were autosomal recessive, 1 patient had DNAH11 and HYDIN gene mutations, and 4 patients had no gene mutations. The median follow-up period was 36.3 (1.9–79) months. All participating patients were applied to myeloablative conditioning (MAC) regimens. The rates of OS, EFS, and GEFS within three years were 87.5%, 85.3%, and 75.2%, respectively. The total graft failure and the total mortality rate were 5.3% and 11.1%. The cumulative incidence of acute GVHD was 53.8% and the incidence of chronic GVHD was 12.9%, The incidence of chronic GVHD was higher for patients who received unrelated donor cord blood stem cell transplantation (UD-CB) (P = 0.001). Chronic GVHD and coinfections are the risk factors for OS and EFS in patients with CGD after receiving allo-HSCT. UD-CB is a risk factor for EFS and the presence of pneumonia before transplantation is a risk factor for OS. In conclusion, through this study, we have demonstrated that allo-HSCT has excellent efficacy in the treatment of CGD in children, especially, RD-haplo is associated with a lower rate of graft failure incidence and mortality than the treatment modalities of other donor type. Therefore, allo-HSCT is strongly recommended when a well-matched donor is available. If a well-matched donor is not available, the HLA-mismatched donor should be carefully evaluated, and the conditioning regimen modified accordingly.