Use Of Cellular Therapy Research Articles

The role of intensive care nurses in cellular treatments during the COVID-19 pandemic.

Today, the use of cellular therapies as an effective treatment in the field of health is increasing. In the COVID-19 pandemic or similar situations, cellular therapies may be sometimes life-saving. The COVID-19 pandemic has shown us that the training of intensive care nurses in special cases, such as cellular therapies, is insufficient. The study aimed to determine the duties, responsibilities and training of intensive care nurses on mesenchymal stem cells (MSCs) transplantation to critically ill patients during the COVID-19 pandemic. This descriptive and retrospective study was conducted on 107 critically ill patients diagnosed with COVID-19 infection and followed up in the intensive care unit (ICU) between April 2020 and April 2022. Each patient was transplanted MSCs by intravenous infusion three times. Before starting cellular therapy applications, intensive care nurses were selected to work on this treatment modality. Each nurse was given theoretical and practical training by experienced instructors. Intensive care nurses trained for MSCs transplants took part in the pre-application, preparation, application and post-application period. MSCs were checked by the ICU nurses in the pre-application period. Patients' vital signs, existing catheters, consciousness status and parameters were checked by nurses in the preparation and application period. No side effects and complications were observed in patients during MSCs transplantation and within the first 24 h. Patients' late complications and mortality were recorded by nurses during the post-application periods. We recommend that nurses working especially in Level 3 ICUs receive training and certification in cellular therapies, especially in hospitals where advanced/cellular treatments are applied. Intensive care nurses are actively involved in every phase of the application of MSCs. Especially before such special practices, which came to the fore with the COVID-19 pandemic, training should be organized for intensive care nurses.

A robust quality infrastructure is key to safe and effective delivery of immune effector cells: how FACT-finding can help

AbstractImmune effector cells (IECs) include a broad range of immune cells capable of modulating several disease states, including malignant and nonmalignant conditions. The growth in the use of IECs as both investigational and commercially available products requires medical institutions to develop workflows/processes to safely implement and deliver transformative therapy. Adding to the complexity of this therapy are the variety of targets, diseases, sources, and unique toxicities that a patient experiences following IEC therapy. For over 25 years, the Foundation for the Accreditation of Cellular Therapy (FACT) has established a standard for the use of cellular therapy, initially with hematopoietic cell transplantation (HCT), and more recently, with the development of standards to encompass IEC products such as chimeric antigen receptor (CAR)-T cells. To date, IEC therapy has challenged the bandwidth and infrastructure of the institutions offering this therapy. To address these challenges, FACT has established a programmatic framework to improve the delivery of IEC therapy. In this study, we outline the current state of IEC program development, accreditation, and solutions to the challenges that programs face as they expand their application to novel IEC therapy.

Recent advances in the combination of cellular therapy with stem cells and nanoparticles after a spinal cord injury.

Currently, combined therapies could help to reduce long-term sequelae of spinal cord injury (SCI); stem cell therapy at the site of injury in combination with other therapies has shown very promising results that can be transferred to the clinical field. Nanoparticles (NPs) are versatile technologies with applications to medical research for treatments of SCI since they could deliver therapeutic molecules to the target tissue and may help to reduce the side effects of non-targeted therapies. This article's purpose is to analyze and concisely describe the diverse cellular therapies in combination with NPs and their regenerative effect after SCI. We reviewed the literature related to combinatory therapy for motor impairment following SCI that has been published by Web of Science, Scopus, EBSCO host, and PubMed databases. The research covers the databases from 2001 to December 2022. Animal models of SCI have shown that the combination of NPs plus stem cells has a positive impact on neuroprotection and neuroregeneration. Further research is required to better understand the effects and benefits of SCI on a clinical level; therefore, it is necessary to find and select the most effective molecules that are capable of exacerbating the neurorestorative effects of the different stem cells and then try them out on patients after SCI. On the other hand, we consider that synthetic polymers such as poly [lactic-co-glycolic acid] (PLGA) could be a candidate for the design of the first therapeutic strategy that combines NPs with stem cells in patients with SCI. The reasons for the selection are that PLGA has shown important advantages over other NPs, such as being biodegradable, having low toxicity levels, and high biocompatibility; In addition, researchers could control the release time and the biodegradation kinetics, and most importantly, it could be used as NMs on other clinical pathologies (12 studies on www.clinicaltrials.gov) and has been approved by the Federal Food, Drug, and Cosmetic Act (FDA). The use of cellular therapy and NPs may be a worthwhile alternative for SCI therapy; however, it is expected that the data obtained from interventions after SCI reflect an important variability of molecules combined with NPs. Therefore, it is necessary to properly define the limits of this research to be able to continue to work on the same line. Consequently, the selection of a specific therapeutic molecule and type of NPs plus stem cells are crucial to evaluate its application in clinical trials.

Post-Hematopoietic Cell Transplantation Relapsed Acute Lymphoblastic Leukemia: Current Challenges and Future Directions.

Allogeneic hematopoietic cell transplantation (allo-HCT) represents an important and potentially curative treatment option for adult patients with acute lymphoblastic leukemia. Relapse continues to remain the most important factor influencing overall survival post allo-HCT. We discuss early identification, clinical manifestations, and management of relapsed disease. Routine evaluation of measurable residual disease (MRD) and change in donor chimerism play a crucial role in early detection. Pivotal clinical trials have led to FDA approval of multiple novel agents like blinatumomab and inotuzumab. Combining targeted therapy with cellular immunotherapy serves as the backbone for prolonging overall survival in these patients. Donor lymphocyte infusions have traditionally been used in relapsed disease with suboptimal outcomes. This review provides insight into use of cellular therapy in MRD positivity and decreasing donor chimerism. It also discusses various modalities of combining cellular therapy with novel agents and discussing the impact of chimeric antigen receptor T-cell therapy in the setting of post allo-HCT relapse both as consolidative therapy and as a bridge to second transplant.

Activated Mesenchymal Stromal Cell Therapy for Treatment of Multi-Drug Resistant Bacterial Infections in Dogs.

New and creative approaches are required to treat chronic infections caused by increasingly drug-resistant strains of bacteria. One strategy is the use of cellular therapy employing mesenchymal stromal cells (MSC) to kill bacteria directly and to also activate effective host immunity to infection. We demonstrated previously that activated MSC delivered systemically could be used effectively together with antibiotic therapy to clear chronic biofilm infections in rodent models. Therefore, we sought in the current studies to gain new insights into the antimicrobial properties of activated canine MSC and to evaluate their effectiveness as a novel cellular therapy for treatment of naturally-occurring drug resistant infections in dogs. These studies revealed that canine MSC produce and secrete antimicrobial peptides that synergize with most classes of common antibiotics to trigger rapid bactericidal activity. In addition, activated canine MSC migrated more efficiently to inflammatory stimuli, and secreted factors associated with wound healing and fibroblast proliferation and recruitment of activated neutrophils. Macrophages incubated with conditioned medium from activated MSC developed significantly enhanced bactericidal activity. Clinical studies in dogs with chronic multidrug resistant infections treated by repeated i.v. delivery of activated, allogeneic MSC demonstrated significant clinical benefit, including infection clearance and healing of infected tissues. Taken together, the results of these studies provide new insights into antimicrobial activity of canine MSC, and their potential clinical utility for management of chronic, drug-resistant infections.

Abstract LB101: Novel Drug-responsive domain (DRD)-based regulation technology enables tightly controlled activity of potent membrane-bound IL12 in adoptive cell therapies

Abstract Interleukin 12 (IL12) is an attractive cancer immunotherapeutic known to be extremely potent against solid tumors preclinically. However, the clinical utility of IL12 has been limited by toxicities stemming from high systemic cytokine exposure. Thus, armoring cellular therapies such as chimeric antigen receptor T cells (CAR-Ts) or tumor infiltrating lymphocytes (TILs) with IL12 will require technologies that provide tight control of IL12 expression and localization. Herein, we describe a tightly regulated version of IL12 for use in cellular therapies. First, we show that tethering IL12 to the membrane increases the activity of IL12 at the tumor site in vivo while reducing potential systemic toxicities. Membrane bound (mbIL12) was compared to secreted IL12 in the syngeneic CD8 gp100 TCR transgenic PMEL model. In this model, PMEL T cells transduced with mbIL12 demonstrated similar reduction in B16 tumor outgrowth as secreted IL12. Like secreted IL12, mbIL12 enhanced the expansion of PMEL T cells and retained T cell extrinsic activities, such as activation of myeloid cells and remodeling the tumor microenvironment. However, mbIL12 showed reduced toxicity signals as compared to secreted IL12, including a reduction in serum IFNy. Likewise, in a xenograft system, CD19-CARTs expressing human mbIL12 demonstrated enhanced potency against Raji tumors. To enhance the safety of mbIL12 further we sought to regulate its expression using Obsidian’s cytoDRIVE® technology. This platform consists of small, fully human protein sequences called drug responsive domains (DRD)s, such as carbonic anhydrase 2, that enable regulation of expression of a fused target protein under the control of FDA-approved, orally bioavailable small molecule ligands, such as, acetazolamide. In the absence of ligand (the “off-state”), the fusion protein is unfolded and degraded. In the presence of ligand (the “on-state”), the DRD is stabilized, allowing for protein expression and function. Thus, the cytoDRiVE® system acts as a titratable and reversible rheostat for on-demand protein activity. While a single DRD can enable some regulation of mbIL12 levels, we found that adding a modulation hub that increases the multiplicity of DRDs greatly improves regulation. Indeed, these modifications enabled off-state levels of mbIL12 that were indistinguishable from untransduced controls in HEK and Jurkat cell lines as well as primary human CD19 CAR-T-cells using flow cytometry. These modifications also enhanced mbIL12 regulation, leading up to a 30-fold dynamic range of mbIL12 abundance. Regulated mbIL12 was demonstrated to be active by phosphorylation of STAT4 in bystander NK cells in vitro. Combining DRDs with modulation hubs and membrane tethering enables the control of highly potent, previously clinically intractable cytokines, such as IL12, for use in enhancing cell therapies. Citation Format: Sean Gregory Smith, James A. Storer, Dexue Sun, Dan Jun Li, Benjamin Primack, Theresa Ross, Scott LaJoie, Jeremy Tchaicha, Dhruv Sethi, Jan ter Meulen, Michelle Ols. Novel Drug-responsive domain (DRD)-based regulation technology enables tightly controlled activity of potent membrane-bound IL12 in adoptive cell therapies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB101.

Richter's Transformation.

Richter's transformation (RT) occurs when chronic (CLL) transforms into an aggressive lymphoma. Despite improvements in the treatment of CLL, prognosis for RT remains poor. Here, we review current literature of RT, with a focus on novel treatment options. Efforts are underway to improve outcomes for patients with RT. While small molecule inhibitors have limited efficacy as monotherapy, recent developments combining them with chemo-immunotherapy show promise. Studies exploring the use of cellular therapies including chimeric antigen receptor T-cells and bispecific antibodies are ongoing. The current treatment paradigm for RT is to enroll these patients on a clinical trial when available, together with consultation for a consolidative allogeneic stem cell transplant. Trials investigating novel combinations and cellular therapy are ongoing. Determining predictive variables of transformation is imperative to design studies that allow for early identification and intervention for patients with RT.

Safety and Efficacy of Mesenchymal Stromal Cells and Other Cellular Therapeutics in Rheumatic Diseases in 2022: A Review of What We Know So Far.

Although a number of new immunosuppressive agents and biologics have been approved for treating various autoimmune inflammatory rheumatic diseases, there remains a substantial number of patients who have no clinical response or limited clinical response to these available treatments. Use of cellular therapies is a novel approach for the treatment of autoimmune inflammatory rheumatic diseases, with perhaps enhanced efficacy and less toxicity than current therapies. Autologous hematopoietic stem cell transplants were the first foray into cellular therapies, with proven efficacy in scleroderma and multiple sclerosis. Newer, yet unproven, cellular therapies include allogeneic mesenchymal stromal cells, which have been shown to be effective in graft-versus-host disease and in healing Crohn's fistulas. Chimeric antigen receptor T cells are effective in various malignancies, with possible application in rheumatic diseases, as shown in preclinical studies in murine lupus and recently in human lupus. Treg cells are one of the master controllers of the immune response and are decreased in number and/or effectiveness in specific autoimmune diseases. Expansion of autologous Treg cells is an attractive approach to controlling autoimmunity. There are a number of other regulatory cells in the immune system, including Breg cells, dendritic cells, macrophages, and other T cell types, that are in early stages of development as treatments. In this review, the current evidence for the efficacy and mechanisms of actions of cellular therapies already in use or in clinical trials in human autoimmune diseases will be discussed, including the limitations of these therapies and potential side effects.

Cellular Therapy Advances in Chronic Lymphocytic Leukemia and Richter's Syndrome

Over the past 10 years, there have been great treatment advances for chronic lymphocytic leukemia (CLL) with the development of small molecule inhibitors. However, there remains an area of unmet need for patients who progress on novel therapies. The development of cellular therapies in CLL has been hindered by CLL induced immunosuppression. Fortunately, recent progress in various methods in immunomodulation may help overcome this limitation in CLL. These advances have spurred ongoing interest in the development of cellular therapies for CLL, including chimeric antigen receptor (CAR) T cell therapies, bi-specific antibodies, and use of natural killer cells. These novel treatment modalities may hold promise for patients with refractory, and potentially transformed disease. Here, we discuss the development of CAR-T cell therapy in CLL and the impact of combining CAR-T and small molecule inhibitors on treatment outcomes, the evolving role of bi-specific antibodies and natural killer cells, and comment on the use of cellular therapies for Richter's syndrome.

Is Culture Expansion Necessary in Autologous Mesenchymal Stromal Cell Therapy to Obtain Superior Results in the Management of Knee Osteoarthritis?-Meta-Analysis of Randomized Controlled Trials.

Study Design: Meta-analysis. Objectives: We aimed to analyze the impact of cultured expansion of autologous mesenchymal stromal cells (MSCs) in the management of osteoarthritis of the knee from randomized controlled trials (RCTs) available in the literature. Materials and Methods: We conducted independent and duplicate electronic database searches including PubMed, Embase, Web of Science, and Cochrane Library until August 2021 for RCTs analyzing the efficacy and safety of culture-expanded compared to non-cultured autologous MSCs in the management of knee osteoarthritis. The Visual Analog Score (VAS) for pain, Western Ontario McMaster University’s Osteoarthritis Index (WOMAC), Lysholm score, Knee Osteoarthritis Outcome Score (KOOS), and adverse events were the analyzed outcomes. Analysis was performed in R-platform using OpenMeta [Analyst] software. Results: Overall, 17 studies involving 767 patients were included for analysis. None of the studies made a direct comparison of the culture expanded and non-cultured MSCs, hence we pooled the results of all the included studies of non-cultured and cultured types of MSC sources and made a comparative analysis of the outcomes. At six months, culture expanded MSCs showed significantly better improvement (p < 0.001) in VAS outcome. Uncultured MSCs, on the other hand, demonstrated significant VAS improvement in the long term (12 months) in VAS (p < 0.001), WOMAC (p = 0.025), KOOS score (p = 0.016) where cultured-expanded MSCs failed to demonstrate a significant change. Culturing of MSCs did not significantly increase the complications noted (p = 0.485). On sub-group analysis, adipose-derived uncultured MSCs outperformed culture-expanded MSCs at both short term (six months) and long term (12 months) in functional outcome parameters such as WOMAC (p < 0.001, p = 0.025), Lysholm (p < 0.006), and KOOS (p < 0.003) scores, respectively, compared to their controls. Conclusions: We identified a void in literature evaluating the impact of culture expansion of MSCs for use in knee osteoarthritis. Our indirect analysis of literature showed that culture expansion of autologous MSCs is not a necessary factor to obtain superior results in the management of knee osteoarthritis. Moreover, while using uncultured autologous MSCs, we recommend MSCs of adipose origin to obtain superior functional outcomes. However, we urge future trials of sufficient quality to validate our findings to arrive at a consensus on the need for culture expansion of MSCs for use in cellular therapy of knee osteoarthritis.

Preliminary phenotypic characterization of hematopoietic and non-hematopoietic stem cells from stimulated bone marrow without expansion for its use in cellular therapy

Preliminary phenotypic characterization of hematopoietic and non-hematopoietic stem cells from stimulated bone marrow without expansion for its use in cellular therapy

Isolation and characterization of exosomes from adipose tissue-derived mesenchymal stem cells.

Mesenchymal stem cells (MSCs) are the subject of intense research as they are a potential therapeutic tool for several clinical applications. The new MSCs action models are focused on the use of MSC-derived secretome which contains several growth factors, cytokines, microRNAs, and extracellular vesicles such as exosomes. Exosomes have recently emerged as a component with great potential involved as mediators in cellular communication. The isolation and identification of exosomes has made it possible for them to be used in cell-free therapies. The purposes of this study are: (i) to detect exosomes released into adipose-derived MSC conditioned cell culture medium, (ii) to identify exosome morphology, and (iii) to carry out a complete characterization of said exosomes. Moreover, it is aimed at determining which method for exosome isolation would be best to use. Precipitation has been identified as a highly useful method of exosome isolation since it provides higher efficiency and purity values than other methods. A broad characterization of the exosomes present in the MSC-conditioned medium was also carried out. This work fills a gap in the existing literature on bioactive molecules which have attracted a great deal of interest due to their potential use in cellular therapies.

Distinct Transcriptional Signatures Distinguish the Emergence of Multipotent Progenitors and Hematopoietic Stem Cells from Endothelial Precursors in the Murine Embryo

During embryonic development, blood cells emerge from hemogenic endothelium (HE), producing waves of hematopoietic progenitors prior to the emergence of rare hematopoietic stem cells (HSCs), which have the unique ability to self-renew and generate all cell types of the adult hematopoietic system. HSCs have significant potential for use in cellular therapies and disease modeling. However, efforts to generate HSCs in vitro from pluripotent stem cells (PSCs) have been limited by an incomplete understanding of the unique phenotypic markers and transcriptional programs that distinguish HE with HSC potential. Previous studies have demonstrated that yolk sac-derived erythromyeloid progenitors and HSCs originate from distinct populations of HE. However, it is not known whether the earliest lymphoid-competent progenitors, multipotent progenitors, and HSCs originate from HE with common phenotypic and transcriptional properties. To investigate this, we combined index sorting of single hemogenic precursors with stromal co-culture that enables simultaneous detection of HSC and multilineage hematopoietic potential, to functionally validate surface markers that may distinguish hemogenic precursors with different hematopoietic fates. We previously found that the co-expression of two markers, CD61 and EPCR, identifies a subset of VE-Cadherin+ endothelial cells from the mouse P-Sp/AGM region (para-aortic splanchnopleura/aorta-gonad-mesonephros, where the first HSCs are generated from HE between E9 and E11 in development) enriched phenotypically for arterial endothelial surface markers (e.g. Dll4, CD44) and functionally for hemogenic precursors with HSC potential. However, this population remains heterogeneous, containing clonal hemogenic precursors with the potential for HSC as well as multilineage progenitor-restricted fates. Here, we report that expression of arterial marker CXCR4 further enriched for functional HSC potential in hemogenic precursors in the P-Sp/AGM between E9 and E10, when the first clonal HSC precursors are detected at rare frequency. In contrast, we detected more abundant clonal HE with multilineage hematopoietic potential (producing lymphoid, erythroid, and myeloid progeny in vitro but lacking HSC potential) at the same stage, which are distinguished by comparatively lower CXCR4 expression. To investigate transcriptional differences between HE populations differentially expressing CXCR4, we performed single-cell RNA sequencing of E9 P-Sp-derived VE-Cadherin+CD61+EPCR+ cells. Using an unbiased gene module analysis based on graph autocorrelation in the Monocle 3 platform to identify genes that co-vary over pseudotime, we found that Cxcr4 is uniquely expressed in a subset of cells simultaneously enriched for arterial-specific genes (including Dll4, Efnb2, Hey2, Sox17, Cd44) and genes with established roles in HSC maintenance and self-renewal (including Mecom, Cdkn1c, H19, Txnip, Kmt2a). Conversely, expression of these genes is decreased in cells undergoing the endothelial to hematopoietic transition at this stage based on pseudotemporal ordering, concomitant with increasing expression of hematopoietic-specifying transcription factors Runx1 and Gfi1, and other genes associated with definitive hematopoiesis (egs. Myb, Kit, Hlf, Gata2, Mpl, Lyl1). We also examined the aggregate expression of established HSC-specific signature genes from published data sets across pseudotime, and found that they exhibit similar expression dynamics to that of Cxcr4 and Dll4, reaching peak expression prior to the initiation of Runx1 and Gfi1 expression. Altogether, our studies support a model in which the initial populations of multipotent progenitors and HSCs emerge independently from HE in the P-Sp/AGM. Furthermore, our findings suggest that HE with HSC competence is uniquely defined by co-expression of arterial endothelial genes and genes encoding HSC self-renewal factors, providing insight into the earliest transcriptional programs that must be coordinated to drive HSC fate from endothelial precursors. Future studies will focus on identifying the signal pathways whose integration promotes expression of these HSC-defining transcriptional programs in endothelial cells, with the goal of advancing methods for HSC generation in vitro. Disclosures Bernstein: Lyell Immunopharma: Current equity holder in publicly-traded company, Other: Laboratory Support; Deverra Therapeutics: Current equity holder in publicly-traded company.

Implementation of experimental cellular (cellular-genetic) therapies on the example of eye diseases

The development of technology and a modern research approach in the 21st century has enabled access to new therapies in many fields of medicine and nanotechnology. Work on the use of cellular therapies is carried out in leading centers around the world. According to the "clinicaltraial.gov" service, it is estimated that there are about 5.5 thousand clinical trials using stem cells. The classification of stem cells is based on their potential to differentiate into other cells, tissues, organs or the whole organism. In cell therapy 3 groups of stem cells are used: Pluripotent Stem Cells (PSC), multipotent and unipotent, which have two common features: the ability to self-revalue, that is, to divide and to differentiate in many directions. To date, there are no objective, randomized clinical trials that would clearly determine the efficacy and safety of the cell therapy used in ophthalmic diseases. Hope is given by gene therapies such as the recently approved Luxturna™ gene therapy used in hereditary retinal degeneration caused by mutations in the RPE65 gene. Research is currently underway on experimental cell therapies to treat the following diseases of the optic system: glaucoma, retinopathy, age-related macular degeneration (AMD), optic nerve atrophy, retinal pigmentation (RP) and Stargardt's disease. Stem cell-based medical therapies are a promising and rapidly developing method of innovative treatment, especially for conditions that were previously considered incurable. The use of experimental cellular gene therapies in diseases of the visual organ gives hope to both patients and scientists, but the age of regenerative medicine has yet to come.

213. Study comparing NTG-101, a growth factor-based therapy with mesenchymal stem cell (MSC) based treatment of degenerative disc disease (DDD) in pre-clinical rodent model

BACKGROUND CONTEXT Currently there is significant interest in the use of cellular therapies (such as stem cells) to treat degenerative disc disease (DDD). We recently explored the potential use of bone marrow derived stem cells (BMSCs) and cartilage-derived stem cells (CDSCs) in a small animal model of DDD as compared to our recently developed molecular therapy ‘NTG-101’. PURPOSE To compare the therapeutic efficacy of NTG-101 with BMSCs or CDSCs to treat DDD in rodent model. STUDY DESIGN/SETTING In vivo laboratory experimentation involving female Wistar rats aged 12-weeks. PATIENT SAMPLE No human subjects. Animal Care approved study involving female Wistar rats using validated rat-tail injury and injection model. OUTCOME MEASURES Histology, immunohistochemistry, Western blotting, gene expression. METHODS CDSCs were derived from the tibial and femoral plateaus of Wistar rats and sphere forming cells were passaged further in serum free, stem cell specific media. BMSCs were obtained from the femurs of these animals and cultured as adherent cells in media containing 8% fetal bovine serum (FBS) following standard protocols. Stemness was confirmed by differentiation into multiple lineages. In parallel, we induced DDD in caudal intervertebral discs (IVDs) of 12 weeks old Wistar rats (N=30) using image-guided needle puncture injury. Ten weeks later, animals were randomized in six treatment groups (n=5 animals/group) and received an intradiscal injection (8µl) containing:G1-phosphate buffered saline (PBS, 1X, pH=7.2); G2-NTG-101(1X);G3-CDSCs (150,000 cells/disc) suspended in PBS; G4-BMSCs (150,000 cells/disc) suspended in PBS;G5-CDSCs (150,000 cells/disc) suspended in NTG-101(1X); G6-BMSCs (150,000 cells/disc) suspended in NTG-101(1X). At the end of experiment (ie, 10 weeks post-injection, 20 weeks post-injury), animals were humanely euthanized and IVDs were harvested for histology, immunohistochemistry and Western blotting. RESULTS Gene expression analysis using quantitative real time PCR confirmed expression of pluripotency (Oct4, Nanog, Sox2) and mesenchymal stem cell surface markers (CD29, CD44, CD90, CD105, CD133, CD166) in all stem cells. CDSC and BMSCs differentiation into chondrogenic, osteogenic and adipogenic lineages was confirmed, thereby validating their stemness potential. Discs injected with PBS developed a degenerative phenotype with the loss of NP morphology, cellularity and the development of a degenerative fibrocartilaginous phenotype. On the other hand, IVDs injected with NTG-101 or CDSCs alone showed a significant recovery. The morphology and cellularity of discs injected with BMSCs alone or in combination with NTG-101 was not significantly different from those discs injected with PBS. NTG-101 injected discs showed a healthy NP rich in notochordal and resident stem cells as revealed by nuclear Brachyury and Oct4 respectively, whereas discs injected with stem cells alone or stem cells plus NTG-101 did not. CONCLUSIONS Our preclinical data demonstrated that single intradiscal injection of NTG-101 is a superior and effective therapy for regeneration of IVD-NP in comparison to mesenchymal stem cell-based therapies for treatment of DDD. FDA DEVICE/DRUG STATUS This abstract does not discuss or include any applicable devices or drugs.

To Infinity and Beyond: Developing a Nursing Cellular Therapy Program of Care

Topic Significance & Study Purpose/Background/Rationale For nurses to be proficient in caring for patients with cellular treatment protocols, the need for a formal, standardized educational program and practice standards for nursing care is critical. The use of cellular therapy is growing beyond standard of care for malignancy and hematologic disease. Cellular therapy administration in multiple clinical care settings makes it difficult to ensure all nurses have the same level of knowledge and practice which could result in gaps of care for patients. Researchers are actively investigating adoptive cell transfer including Tumor Infiltrating Cells, Dendritic Cells, Chimeric Antigen Receptor T-Cells, as well as other cellular therapies. The need for a formalized Cellular Therapy Program of Care was identified to standardize education and practice for nurses within the oncology, transplant, medical surgical, ambulatory, pediatric, and intensive care specialties. Methods, Intervention, & Analysis Trends in approved therapies as well as new research protocols show an increase in use of cellular therapies. Additionally, application of cellular therapy has expanded to diseases other than cancer and encompasses multiple routes of administration, e.g. intradermal, intraperitoneal, and intrathecal. Nursing programs of care that have not traditionally administered cellular products may begin to implement these treatments. Findings & Interpretation A multi-disciplinary team developed a standardized approach for education and practice that includes; education in cellular science and chemotherapy, standards of practice and procedures on cellular administration, training in complications of cellular medicine, competency assessment, and standardized documentation practices Discussion & Implications The Nursing Cellular Therapy Program of Care is in its early stages and outcome evaluation will be measured through patient quality indicators, documentation compliance, and ongoing competency evaluation. Formalization of a cellular program standardizes nursing practice and ensures patient safety.

Outcomes after Allogeneic Hematopoietic Stem Cell Transplantation for Pediatric Acute Myeloid Leukemia in the Contemporary Era

High risk or relapsed pediatric acute myeloid leukemia (AML) is potentially curable with allogeneic hematopoietic stem cell transplantation (HSCT). Significant improvements have been made with supportive care practices, but there are limited HSCT outcome data for pediatric AML patients in the contemporary era. We report retrospective data in 87 pediatric patients with high risk or relapsed AML, who underwent HSCT at a single institution between 2008 and 2018. The median age at time of HSCT was 8 years (range: 0.6-20 years). Forty three patients (49%) were transplanted in CR1, 18 (21%) in CR2, and 26 (30%) with active disease (defined as any evidence of disease at the time of HSCT). Patients were transplanted using matched related (MRD, n=21), matched unrelated (MUD, n=25), mismatched unrelated (MMUD, n=15), haplo-identical (haplo, n=12) or umbilical cord blood (UCB, n=14) grafts. The majority of patients (78/87, 90%) received myeloablative conditioning. MRD and UCB graft recipients did not receive serotherapy. All other graft recipients (n=52) received serotherapy, the majority using an alemtuzumab-based conditioning regimen, 42/52 (81%). Haplo-identical transplants were performed using CD34+ selected grafts. Three-year overall survival (OS) and disease free survival (DFS) for the entire cohort were 52% (95% CI: 41-62%) and 49% (95% CI: 38-59%) respectively. OS differed significantly according to disease status at time of transplant (p=0.018), with 3 year OS: 60%, (95% CI: 43-74%) for those in CR1, 56%, (95% CI: 31-75%) for those in CR2 and 34% (95% CI: 17-52%) for those with active disease. OS differed by donor type (p=0.058). OS was comparable for patients with MRD (71%, 95% CI: 46-86%) and 10/10 MUD (59%, 95% CI: 37-75%) (p=0.67). Patients with alternate donor sources had worse outcomes (MMUD 33%, 95% CI: 10-59%; UCB 43%, 95% CI: 18-66%; Haplo 33%, 95% CI: 10-59%) as compared to MRD (MMUD vs MRD, p=0.042, UCB vs MRD, p=0.046, haplo vs MRD p=0.017). There was a low incidence of Grade III-IV acute GVHD (8%, n=7) and extensive chronic GVHD (9%, n=7) likely secondary to the use of alemtuzumab-based conditioning regimens. The 3-year non-relapse mortality was low at 11% (95% CI: 5-19%). The primary cause of death was relapse, with a 3-year cumulative incidence of relapse of 40%, (95% CI: 30-50%). Patients with active disease at HSCT had significantly higher cumulative incidence of relapse (sHR: 2.42, 95% CI: 1.18-4.95, p=0.016) compared to patients in remission. In the contemporary era, outcomes after allogeneic HSCT are comparable for MRD and 10/10 MUD for pediatric AML. Myeloablative and alemtuzumab based conditioning regimens were well tolerated with low rates of NRM and GVHD. Relapse remains the major cause of treatment failure. Future trials evaluating use of cellular therapies and targeted agents post-HSCT will be needed.

Tendinopatías: etiología, histopatología y avances terap

&#x0D; Objetivos: Describir los principales cambios histopatológicos presentes en las tendinopatías, su patogénesis y los avances terapéuticos encontrados en la literatura. &#x0D; Palabras Claves: Tendinopatías, histología, patogénesis, inflamación, tendinosis, regeneración. &#x0D; Materiales y métodos: Se realizó una búsqueda de literatura relacionada con las tendinopatías su patogénesis, cambios histológicos, avances terapéuticos en el uso de células madres, factores de crecimiento (plasma rico en plaquetas) y terapia farmacológica. Se utilizaron las bases de datos de pubmed, clinical key, science direct. Se incluyeron artículos de revisiones bibliográficas entre 2005 al 2018, escritos en inglés. &#x0D; Conclusiones: Las tendinopatías tienen un complejo origen multifactorial, donde los factores extrínsecos e intrínsecos juegan un papel importante para el desarrollo de dicha afectación. La inflamación tiene un rol determinante en el desarrollo de la patogenia de las tendinopatías. La terapia farmacológica antiinflamatoria tiene efectos a corto plazo en la disminución del dolor, sin embargo, resulta contraproducente a largo plazo. Existe controversia sobre el uso del Plasma rico en plaquetas (PRP) y de terapia celular (stem cells), se requiere de mayor investigación que pueda evidenciar la eficacia de dichos tratamientos.

Outcomes after Allogeneic Hematopoietic Stem Cell Transplantation for Pediatric Acute Myeloid Leukemia in the Contemporary Era

High risk or relapsed pediatric acute myeloid leukemia (AML) is potentially curable with allogeneic hematopoietic stem cell transplantation (HSCT). The use of a matched related donor (MRD) is the standard of care for these patients, but in the majority of patients a MRD is not available. Significant improvements have been made with supportive care practices, but there are limited outcome data for AML in pediatric patients transplanted in the contemporary era. We report retrospective results of HSCT in eighty-seven pediatric patients with high risk or relapsed AML performed at a single institution between 2008 and 2018. The median age at the time of HSCT was 8 years old (range: 0.6-20 years). More than 2/3rd of patients were in remission at time of HSCT with 43 patients (49%) transplanted in CR1 and 18 patients (21%) in CR2. Twenty six patients (30%) had active disease (defined as any evidence of disease) at the time of HSCT. Patients were transplanted using a matched related donor (MRD, n=21), a matched unrelated donor (MUD, n=25), a mismatched unrelated donor (MMUD, n=15), a haplo-identical donor (haplo, n=12) or an umbilical cord blood (UCB, n=14) graft. The majority of patients (78/87, 90%) received myeloablative conditioning while 9 patients (10%) received reduced intensity conditioning regimens. MRD and UCB graft recipients did not receive serotherapy. MUD, MMUD and haplo graft recipients (n=52) received serotherapy, the majority of whom received an alemtuzumab-based regimen, 42/52 (81%). Haplo-identical transplants were performed using CD34+ selected grafts. Three-year overall survival (OS) and disease free survival (DFS) for the entire cohort were 52% (95% CI: 41-62%) and 49% (95% CI: 38-59%) respectively. OS differed significantly according to disease status at time of transplant (p=0.018), with 3 year OS: 60%, (95% CI: 43-74%) for patients in CR1, 56%, (95% CI: 31-75%) for patients in CR2 and 34%, (95% CI: 17-52%) for patients with active disease. OS differed by donor type (p=0.058). OS was comparable for patients with MRD (71%, 95% CI: 46-86%) and 10/10 MUD (59%, 95% CI: 37-75%) (p=0.67). Patients with alternate donor sources had worse outcomes (MMUD 33%, 95% CI: 10-59%; UCB 43%, 95% CI: 18-66%; Haplo 33%, 95% CI: 10-59%) as compared to MRD (MMUD vs MRD, p=0.042, UCB vs MRD, p=0.046, haplo vs MRD p=0.017). There was a low incidence of Grade III-IV GVHD (8%, n=9) and extensive chronic GVHD (9%, n=7) likely secondary to the use of alemtuzumab-based conditioning regimens for MUD, MMUD and haplo donors. The 3-year non-relapse mortality was low at 11%, (95% CI: 5-19%). The primary cause of death was relapse, with a 3-year cumulative incidence of relapse of 40%, (95% CI: 30-50%). Patients with active disease at transplant had significantly higher cumulative incidence of relapse (sHR: 2.42, 95% CI: 1.18-4.95, p=0.016) compared to patients in remission. There was no difference in cumulative incidence of relapse between patients in CR1 and CR2 (p=0.598). In the contemporary era, outcomes after allogeneic HSCT are comparable for MRD and 10/10 MUD for pediatric AML. Myeloablative conditioning regimens that incorporated alemtuzumab were well tolerated with low rates of NRM and very low rates of acute and chronic GVHD. Relapse remains the major cause of treatment failure and future trials evaluating use of cellular therapies and targeted agents post-HSCT will be needed to improve current rates of relapse. Disclosures No relevant conflicts of interest to declare.

The Effect of Early Rounds of ex vivo Expansion and Cryopreservation on the Adipogenic Differentiation Capacity of Adipose-Derived Stromal/Stem Cells

Multipotent adipose-derived stromal/stem cells (ASCs) are candidates for use in cellular therapies for the treatment of a variety of conditions/diseases. Ex vivo expansion of freshly isolated ASCs may be necessary prior to clinical application to ensure that clinically relevant cell numbers are administered during treatment. In addition, cryopreserving cells at early passages allows for storage of freshly isolated cells for extended periods of time before expanding these cells for clinical usage. There are however several concerns that these laboratory-based procedures may alter the characteristics of the cells and in so doing decrease their regenerative potential. In this study we report on the impact of early rounds of cryopreservation (P0) and ex vivo expansion (P0 to P5) on the phenotypic characteristics and adipogenic differentiation potential of ASCs. Our results show that ASCs that upregulate CD36 expression during adipogenic differentiation gradually decrease with increasing expansion rounds. The consequent decrease in adipogenic differentiation capacity was evident in both gene expression and flow cytometry-based phenotypic studies. Successive rounds of expansion did not however alter cell surface marker expression of the cells. We also show that early cryopreservation of ASCs (at P0) does not affect the adipogenic differentiation potential of the cells.