Red Marrow Dosimetry and Stem Cell Reinfusion in High Dose 90Y - Ibritumomab Tiuxetan.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2187-2187 ◽  
Author(s):  
Anna Vanazzi ◽  
Daniele Laszlo ◽  
Marta Cremonesi ◽  
Chiara Maria Grana ◽  
Stefano Papi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Image Analysis ◽  
Activity Concentration ◽  
Activity Level ◽  
Alternative Methods ◽  
Ibritumomab Tiuxetan ◽  
Time Activity ◽  
Bone Marrow Biopsies ◽  
Red Marrow

Abstract BACKGROUND: Red marrow (RM) is the critical organ in Radioimmunotherapy (RIT) with 90Y-ibritumomab tiuxetan, especially at myeloablative activities. Different methods for the evaluation of the adsorbed dose in RM have been investigated. The blood method relates the blood time-activity curve to RM by a factor that accounts for the activity concentration ratio between RM and blood. Alternative dosimetric approaches including image analysis and marrow aspirations (MA) were introduced for RM dose comparison in order to collect information about the dose delivered and to verify the correct time of Autologous Stem Cells Transplantation (ASCT). PATIENTS AND METHODS: Twenty-six patients affected by resistant/refractory B-cell Non Hodgkin Lymphoma (NHL) were enrolled in a phase I/II study to determine the feasibility and safety of High Activity (HA) of 90Y-ibritumomab tiuxetan followed by ASC support. Three 90Y-ibritumomab tiuxetan activity levels were planned: 30 MBq/kg (4 patients), 45 MBq/kg (4 patients), 56 MBq/kg (18 patients). One week before treatment all patients underwent dosimetry with 111Inibritumomab tiuxetan. The activity concentration was measured in 9 serial blood samples up to 7 days post injection (pi) and planar images in all patients, in MA in 8 out of 18 patients planned at 56 MBq/kg (4 patients at 160 h pi; 4 patients on day planned for ASCT, i.e. day 0). ASC were reinfused 13 days after 90Y-ibritumomab tiuxetan. ASCT was considered at low risk when the adsorbed dose to reinfused stem cells (rSC) (i.e. dose to RM at time of ASCT) was < 50 mGy. Engraftment was considered delayed if on day 28 from reinfusion ANC did not reach 1.0x109/L or PLT were not ≥ 20.0x109/L. RESULTS: A median of 4,25x106/kg (1,45–20x106/kg) CD34+ cells were reinfused for each patient. A delay of engraftment occurred in only 1 patient who received 56 MBq/kg with a baseline ANC count of 0.20x109/L. Bone marrow biopsies were negative for disease localization in all patients at baseline, but 10 out of 26 had shown prior lymphoma infiltration. Considering the blood method the adsorbed dose to rSC resulted < 50 mGy in all patients with a median adsorbed dose of 11 mGy (4 – 28 mGy). ASC would be reinfused earlier in all patients except one. In particular, the 1st day with a dose to rSC < 50 mGy ranged from day -9 to day -5 at 30 MBq/kg; from day -8 to day -4 at 45 MBq/kg; from day -9 to day 0 at 56 MBq/kg, with a median value on day -3. We observed a possible correlation between total activity administered, dose delivered to RM and dose detectable on day of reinfusion, with higher doses to RM and rSC for those patients planned to received the highest activity. We also evaluated the dose to rSC if reinfusion would be performed 7 days after RIT: only 6 out of 26 patients showed a dose to SC < 50 mGy (2 at the 1st activity level, 3 at the 2nd and 1 at the 3rd activity level). Considering the MA method, the median adsorbed dose to the rSC resulted 25 mGy (9 - 69 mGy): in 2 out of 8 patients evaluated the adsorbed dose to the rSC was > 50 mGy on day 0 (69 and 51 mGy respectively). Even if ASC would be reinfused earlier in 5 out of 8 patients, the first day with a dose to rSC < 50 mGy ranged from day -3 to day +1. Despite bone marrow biopsy being negative in all patients at baseline, scintigraphic images (L2–L4 uptake) showed spine marrow uptake in almost all patients. CONCLUSIONS: Although alternative methods investigated - MA and imaging - are affected by uncertain factors (blood contamination of MA, difficult L2-L4 uptake image quantification), both indicate comparable results, about 2.5-folds higher, suggesting that the blood method provides an underestimate of RM dose. ASCT performed 13 days after HA 90Y-ibritumomab tiuxetan is generally safe. We suggest caution in anticipating the timing of ASCT with respect to 90Y-ibritumomab tiuxetan in those patients planned to receive an activity > 45 MBq/kg: alternative dosimetric approaches including image analysis and MA should be considered.

Bone Effect of Botezomib in Patients with Relapse and Smoldering Myeloma; A Computer Assisted Image Analysis Study of Bone Marrow Core Biopsies,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3996-3996
Author(s):  
Mohamed E Salama ◽  
Graham E. Wagner ◽  
Tamara Berno ◽  
Jessica Kohan ◽  
Fenghuang Zhan ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Image Analysis ◽  
Bone Marrow Biopsy ◽  
Computer Assisted ◽  
Weekly Dose ◽  
Bone Marrow Biopsies ◽  
Bortezomib Treatment ◽  
Smoldering Myeloma ◽  
Bone Effect

Abstract Abstract 3996 Background: Bone loss and related complications including bone pains, fractures and hypercalcemia are major causes of morbidity and mortality in multiple myeloma (MM) patients. Bone growth/loss (Bone mineral densitometry) can be monitored by dual x-ray absorptiometry (DXA) in patients with smoldering myeloma (SMM). We previously reported a novel quantitative method to asses trabecular volume (TV) using whole scanned slides and image analysis (WSI) obtained from bone marrow biopsy (Teman et al. 2010). This method provides a low cost reproducible mean to assess TV in archival paraffin embedded biopsy materials. Velcade has been shown to produce an anabolic bone effect in relapsed/refractory MM patients and in this study, we examine the effect of low-dose bortezomib (Velcade) in SMM patients using the WSI methodology. Methods: Bone marrow biopsy slides obtained before, during and after bortezomib treatment were used to evaluate TV. H&E stained core biopsy slides were scanned using Scan Scope XT system (Aperio Technologies, Vista, CA) into digital whole slide images that is viewable on Aperio Image Scope. We developed classifier algorithms using Genie (Aperio) pattern recognition image analysis software (PRIA) that were adept at identifying bone, hematopoietic tissue, and clear glass. The calculated bone area (TV) was represented as a ratio of the total hematopoietic area for each biopsy event. Slides were excluded if the analysis available area was less than 6mm2 or could not be classified correctly to the satisfaction of the pathologist Mixed-effects models were used to compare bone TV/hematopoietic ratios (HR) over time and between the different groups, as well as assess any correlation with that ratio and light chain, B-2-microglobulin, and plasma cell levels. Results were considered statistically significant if p<0.05. Results: Slides from 253 consecutive biopsies composed the study materials. 45 were excluded due to significant artifacts or small analysis areas (<6mm2). 208 bone marrow biopsies from 43 patients were included in the analysis. The group included 26 maintenance, 12 relapsed, and 5 smoldering patients; The relapsed and maintenance patients received Bortezomib alone or in combination for a minimum of three cycles; smoldering patients received bortezomib as part of a phase 2 study at the weekly dose of 0.7mg/kg. All maintenance and relapsed, patients had previously received bone marrow transplant with a median 68 years of age 29 were male. Median baseline TV/HR was 32.9%for maintenance 29.8% for relapse and 33.1% smoldering groups. A median increment of TV/RH (17%) was observed after Bortezomib treatment in all groups of patients (p<0.0001). Conclusion: Analysis of bone associated changes after Bortezomib exposure in patients with multiple myeloma by Scan Scope XT demonstrate a post treatment overall gain in bone formation. Monitoring bone indices in patients with multiple myeloma with PRIA may provide a valid tool to assess treatment associated bone effect. Disclosures: No relevant conflicts of interest to declare.

A Phase I Study Of KB004, a Novel Non-Fucosylated humaneered® Antibody, Targeted Against The Receptor Tyrosine Kinase EphA3, In Advanced Hematologic Malignancies

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3838-3838 ◽  
Author(s):  
Jeffrey E Lancet ◽  
Andrew H Wei ◽  
Simon TS Durrant ◽  
Mark S. Hertzberg ◽  
Ronan T. Swords ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Phase 1 ◽  
Hematologic Malignancies ◽  
Clinical Activity ◽  
Sustained Remission ◽  
Eligibility Criteria ◽  
Bone Marrow Biopsies ◽  
Equity Ownership ◽  
Dose Levels

Abstract Background EphA3 is involved in cell positioning in fetal development. In the adult it is an oncofetal antigen re-expressed in hematologic malignancies (blood and bone marrow, leukemic stem cells) and solid tumors (tumor stem cells, neovasculature and stroma) and may be prognostic. KB004 is a humaneered® high affinity antibody targeted against EphA3 with 3 putative mechanisms of action: direct induction of apoptosis in tumor cells, activation of ADCC and disruption of tumor vasculature by induction of endothelial cell rounding and subsequent blood vessel infarction. Herein, we describe results of an ongoing, phase 1 dose-escalation study of KB004 in adult patients with advanced hematologic cancers. Objectives 1) primary: to determine safety and MTD for KB004 in patients with hematologic malignancies refractory to or unfit for chemotherapy; 2) secondary: to examine PK profile, immunogenicity, and clinical activity of KB004. Exploratory objectives include evaluation of EphA3 expression on tumor, stromal, and endothelial cells. Methods This is a multicenter phase 1 study. Key eligibility criteria include: 1) relapsed or refractory hematologic malignancy 2) ECOG PS 0-1; 3) adequate end-organ function. Additional eligibility criteria amended later to protect against hemorrhagic events, included platelets ≥ 10,000/uL (untransfused) and normal coagulation times. A standard 3 + 3 escalation study design (amended to allow up to 6 pts. per cohort in the absence of a DLT) was utilized. KB004 was administered as a 1 or 2 hr infusion on days 1, 8, and 15 of each 21-day cycle, at incremental doses of 20, 40, 70, 100, 140, 190, 250 mg and thereafter 33% increments up to a planned maximum of 700 mg. At 70 mg and above infusion reaction (IR) prophylaxis included an H1 blocker, H2 blocker, acetaminophen and IV steroids. Peripheral blood and bone marrow biopsy specimens for PK analysis and EphA3 expression, respectively, were collected during cycle 1. Results To date, 37 patients (AML 32, MDS 2, lymphoma 1, myelofibrosis 2) received KB004; 20 mg: 9 pts, 40 mg: 3pts, 70 mg: 8 pts, 100 mg: 7pts, 140 mg: 5pts, 190 mg 5pts. At 70 mg, two patients had intracranial hemorrhages 5 and 18 days after last KB004 dose in the context of thrombocytopenia and hyperleukocytosis. A causal relationship to KB004 could not be excluded. One occurred in course 1 and was considered a DLT. KB004 blood levels were near the limit of quantitation at 48 and 96 hours. Following the change to entry criteria no further episodes of serious bleeding or other DLTs or significant changes in soluble clotting factors have been observed. Overall KB004 is well tolerated. The most common toxicity was mild to moderate transient IRs in 28 (76%) patients characterized by chills, elevated temperature, fever, rigors, back pain, nausea, vomiting, hypotension, hypertension, transient hypoxia (in 2 cases). Fourteen % of infusions were slowed due to an adverse event, two (1.2%) were prematurely discontinued but no patient discontinued KB004 secondary to an IR. No other significant KB004-related toxicity has been observed. KB004 Cmax at all dose levels was above the predicted effective concentration (1 ug/ml) and was approximately dose proportional. However at dose levels below 190 mg sustained exposure to cover the entire interval between doses was not achieved. One CRp was observed at the 20mg dose level in a 78 yr-old patient with relapsed AML, who remains on study and in sustained remission for over a year. Serial bone marrow biopsies with KB004 treatment show decreased reticulin and collagen fibrosis. A > 50% reduction in marrow blast percentage was seen in 14% of AML patients, and 59% had overall stable disease beyond 1 cycle. Bone marrow biopsies positive for EphA3 expression with a cut-off of 10% of nucleated cells were obtained in 75% of AML patients. EphA3 was expressed in at least 30% of nucleated cells in the baseline sample of the patient with an ongoing CRp. Conclusion KB004 is a novel agent targeted against EphA3 that is well tolerated with evidence of clinical activity. It is anticipated that 190 mg given over 2 hours will provide sufficient plasma exposure to achieve sustained efficacy over the interval between doses. The study is ongoing. Additional PK, PD, and clinical data will be presented. Disclosures: Durrant: KaloBios: Research Funding. Yarranton:KaloBios: Employment, Equity Ownership; Glaxo: Equity Ownership; EnGen: Equity Ownership, Science Advisor, Science Advisor Other; StemLine Therapeutics: Equity Ownership. Walling:KaloBios: Consultancy; Corcept Therapeutics: Consultancy; Prothena: Consultancy; New Gen Therapeutics: Consultancy; Valent Technologies: Consultancy; LBC Pharmaceuticals: Consultancy; Amgen: Equity Ownership; BioMarin: Equity Ownership; Crown BioScience: Membership on an entity’s Board of Directors or advisory committees.

Stem Cells from New Sources for Cartilage Tissue Engineering

2008 ◽  
Vol 139 (2_suppl) ◽  
pp. P87-P87
Author(s):  
Ulrich R. Goessler ◽  
Jens Stern-Straeter ◽  
Gregor Bran ◽  
Haneen Sadick ◽  
Karl Hoermann ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Adipose Tissue ◽  
Chondrogenic Differentiation ◽  
Cartilage Tissue ◽  
Molecular Characteristics ◽  
Fibronectin Receptor ◽  
Bone Marrow Biopsies ◽  
Adult Mesenchymal Stem Cells ◽  
Signaling Components

Problem The use of adult mesenchymal stem cells (MSC) – especially from new sources including adipose tissue - offers new perspectives in the generation of transplants for reconstructive surgery. The extracellular matrix (ECM) plays a key role in modulating function and phenotype of the embedded cells and contains the integrins as adhesion receptors mediating cell-cell- and cell-matrix-interactions. In our study, characteristic changes in integrin expression during the course of chondrogenic differentiation of MSC from bone marrow and adipose tissue were compared. Methods MSC were isolated from bone marrow biopsies and adipose tissue. During cell culture, chondrogenic differentiation was performed. The expression of Integrins and their signaling components were analyzed with microarray and immunohistochemistry in freshly isolated MSC and after chondrogenic differentiation. Results The Fibronectin-Receptor (Integrin a5b1) was expressed by undifferentiated MSC, expression rose during chondrogenic differentiation in both types of MSC. The components of the Vitronectin/Osteopontin-Receptors (avb5) were not expressed by freshly isolated MSC. Expression rose with ongoing differentiation. Receptors for Collagens (a1b1, a2b1, a3b1) were weakly expressed by undifferentiated MSC and were activated during differentiation. As intracellular signaling components integrin linked kinase (ILK) and CD47 showed increasing expression with ongoing differentiation. For all integrins, no significant differences could be found in the 2 types of MSC. Conclusion Integrin-mediated signaling seems to play an important role in the generation and maintenance of the chondrocytic phenotype during chondrogenic differentiation. The receptors for Fibronectin, Vitronectin, Osteopontin and Collagens in particular might be involved in the generation of the ECM. Intracellularly, their signals might be transduced by ILK and CD47. Significance To fully harness the potential of these cells, future studies should be directed to ascertain their cellular and molecular characteristics for optimal identification, isolation, and expansion.

High Dose 90Yttrium Ibritumomab Tiuxetan with PBSC Support in Refractory-Resistant NHL Patients.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1890-1890 ◽  
Author(s):  
Anna Vanazzi ◽  
Pier Francesco Ferrucci ◽  
Chiara Grana ◽  
Marta Cremonesi ◽  
Margherita Clerici ◽  
...  
Keyword(s):  
Liver Toxicity ◽  
Cell Lymphoma ◽  
Therapeutic Option ◽  
B Cell Lymphoma ◽  
Activity Level ◽  
Hematological Toxicity ◽  
High Dose ◽  
Ibritumomab Tiuxetan ◽  
Organ Doses ◽  
Red Marrow

Abstract We present feasibility and toxicity results of a phase I/II study of High Dose (HD) 90Yttrium (90Y) Ibritumomab Tiuxetan with PBSC support in resistant-refractory NHL patients. From 04/04 to 08/07 25 patients were enrolled. Median age was 68ys (21–75). 21/25 patients had advanced stage disease (III/IV) at diagnosis. 14 patients had Diffuse Large B Cell Lymphoma (DLBCL), 7 Mantle Cell Lymphoma (MCL), 2 Follicular Lymphoma (FL) G3, 1 transformed FL, 1 transformed Marginal Zone Lymphoma (MZL). Median number of prior therapies was 3 (1–6), including Rituximab, RT and HD-CT. Three 90Y-Ibritumomab Tiuxetan activity levels were fixed: 30 MBq/kg, 45 MBq/kg and 56 MBq/kg; 4 patients at the 1st, 4 at the 2nd and 17 at the 3rd level. One week before treatment all patients underwent dosimetry with Indium-111 (111In) ibritumomab-tiuxetan in order to assess organ doses and plan the activity level. PBSC were reinfused 13 days after 90Y-Ibritumomab Tiuxetan. On day 28 from reinfusion engraftment was considered to be delayed if ANC&lt;1.0×109/L or PLT&lt;20.0×109/L. RESULTS: Dosimetry showed acceptable radiation-absorbed doses to normal organs in all cases except one: 1 patient showed an abnormal liver uptake still increasing up to 7 days post injection. The estimated absorbed dose to the liver was 11.3 Gy/GBq, meaning 50 Gy if the prescribed activity of 56 MBq/kg would be administered. Therefore, the patient was excluded from the treatment. The median (24 pts) adsorbed doses for 90Y-Ibritumomab Tiuxetan resulted (mGy/MBq): 0.80 (0.40–1.0), red marrow; 2.1 (1.1–5.4), heart wall; 1.7 (0.3–3.5), lungs; 2.8 (1.8–10.6), liver; 1.9 (0.8–5.0), spleen; 1.7 (0.6–3.8), kidneys; 2.8 (1.3–4.7), testes; 0.5 (0.4–0.8), total-body. Median activity of 90Y-Ibritumomab Tiuxetan delivered: 3.7 GBq, range 2.1–5.55 GBq. All patients engrafted promptly after PBSC transplantation. PLT and ANC-count nadirs were reached 8 and 4 days after transplantation, with median values of 11×109/L PLT (4–35) and 0.01×109/L ANC (0.01–1.09). The time of nadir did not change as a function of 90Y-ibritumomab tiuxetan dose. Median time to engraftment was 12 (0–22) and 20 (1–48) for PLT and ANC respectively. When data were analyzed per dose-level, no statistically significant differences in terms of hematological toxicity were observed. A drop in PLT-counts often occurs after engraftment as a sign of possible late toxicity, particularly in those patients who received more than 3 previous CT regimens. One transient acute G3 liver toxicity was observed at the 3rd level; 1pt died 4 months after treatment due to HCV reactivation; another pt died 2 months after 90Y-Ibritumomab Tiuxetan because of cerebral ischemia; finally we observed a myelodysplastic syndrome in 1pt who had received 45 MBq/kg 2 years after treatment. 22/24 patients are now evaluable for response: 9 CR, 4 PR, 1 SD, 8 PD. CONCLUSION: 90Y-Ibritumomab Tiuxetan at myeloablative activity is feasible with PBSC support and it could be safely delivered also in elderly pts. We suggest dosimetry in order to avoid unpredictable toxicity and an activity of 45 MBq/kg in heavily pretreated patients. Clinical efficacy and mild treatment-related toxicities suggest HD-90Y-Ibritumomab Tiuxetan is an interesting modality treatment to be further investigated as an alternative therapeutic option in ABMT setting.

scholarly journals Mesenchymal Stem Cells from Patients 1 Year Following Autologous Stem Cell Transplantation Have a Pro-Inflammatory and Senescent Phenotype Compromising the Support of Hematopoietic Stem Cells

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4368-4368
Author(s):  
Carin L.E. Hazenberg ◽  
Fiona A.J. van den Heuvel ◽  
Ben N.G. Giepmans ◽  
Jan Jacob Schuringa ◽  
Edo Vellenga
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Stem Cell ◽  
Growth Factor ◽  
Stem Cell Transplantation ◽  
Autologous Stem Cell Transplantation ◽  
Normal Bone Marrow ◽  
Hematopoietic Stem ◽  
Bone Marrow Biopsies ◽  
Normal Bone

Abstract Autologous stem cell transplantation (ASCT) is a frequently applied treatment modality for patients with multiple myeloma (MM) and (relapsing) malignant lymphomas. Normal peripheral blood cell counts are usually observed 1 year post ASCT although the hematopoietic stem cell (HSC) compartment is severely impaired reflected by reduced HSC frequency and quiescence (Haematologica 2013;98:1264). Since HSCs interact intensively with the surrounding microenvironment in the bone marrow and strongly depend on these cells for a proper function, we studied the mesenchymal stem cell (MSC) compartment in patients 1 year post ASCT. We generated a biobank with patient material acquired 1 year after ASCT. Immunohistological studies of bone marrow biopsies post ASCT showed increased expression of CD271 (Nerve Growth Factor Receptor, NGFR) compared to normal bone marrow (NBM, 11.26%±1.2 of bone marrow area versus 1.87%±0.9, p<0.0001) while no difference was observed for the percentage of nestin+ or perivascular CD146+ (Melanoma Cell Adhesion Molecule, MCAM) cells. In addition an increase in CD271+-multilocular adipocytes was noted, reflecting a difference in preferential MSC differentiation. Subsequently MSCs were cultured from the CD34- fraction of the bone marrow mononuclear cells, obtained from post ASCT patients (n=11) and compared to healthy subjects (n=17). MSCs were selected by their plastic-adherency and subsequently replated to generate MSCs. Cultured MSCs from post ASCT and NBM had similar population doubling times (1.92±0.22 and 3.52±1.02 in P4 (passage 4) respectively). In addition no difference in cell surface expression of CD146 and CD271 was demonstrated on MSCs post ASCT as compared to NBM. However, the post ASCT MSCs showed a change in morphology at early passages (P3-4) and premature exhaustion of growth in 45% of the studied patients (n=11) at P5, in contrast to 18% from NBM (n=11). B-galactosidase staining of post ASCT MSCs was increased in P5 and P6 compared to NBM MSCs (20.08%±3.0 vs 9.9%±1.1, p=0.04). To study the functionality of these MSCs, post ASCT MSCs from a low passage (P3 or P4) were used for co-culture experiments with CD34+ cord blood cells in the presence of cytokines SCF, FLT3 and TPO. Co-cultures with MSCs from different post ASCT patients showed a large variation in number of cobblestone-area forming cells (CAFCs, range: 11-163, mean: 81.3±16.0) as well as the size of cobblestone area. This reflects the diversity in HSC support by post ACST MSCs and concurs with the diversity found between patients in the clinical setting. Finally gene profiling performed on cultured post ASCT (n=10) and NBM (n=9) MSCs in early passages (P2 and P3) showed upregulation of proinflammatory genes such as interleukin-6 (IL6) and genes involved in Notch and Transforming Growth Factor-ß (TGF-B) signaling such as Hairy and Enhancer of Split-1 (HES1), and Bone Morphogenetic Protein (BMP)1 and BMP4. These findings were confirmed by quantitative PCR. Foxc1 expression, recently linked to maintenance of hematopoietic stem and progenitor cells, was significantly increased in post ASCT MSCs. Collectively, these data indicate changes in the bone marrow niche, especially in the mesenchymal (CD271+) compartment, inducing premature exhaustion and affecting their supportive role for the HSCs. This damage to the niche may account for the reduced bone marrow reserve observed in patients and generate insight into putative therapeutic targets for improving transplantation strategies. Figure 1a,b,c. Figure 1a,b,c. CD271+ expression is significantly increased in post ASCT bone marrow biopsies (b) compared to normal bone marrow (NBM, a). Quantification of CD271 expression in percentage of total bone marrow area by ImageJ software. * p<0.0001 Figure 2a,b,c,d. Figure 2a,b,c,d. Similar expression of CD146 and CD271 on NBM and post ASCT MSCs. Ns: not significant Premature exhaustion of growth in post ASCT (45%) vs NBM (18%) MSCs before P6 Significant increase in B-galactosidase staining in post ASCT MSCs in P5-P6. * p<0.05 Changed morphology of post ASCT MSCs in vitro, representative example in P4 Disclosures No relevant conflicts of interest to declare.

Saviour-Sibling and the Psychological, Ethical and Judicial Issues that It Creates: Should English and French Legislators Close the Pandora’s Box?

2011 ◽  
Vol 18 (3) ◽  
pp. 293-303 ◽  
Author(s):  
Allane Madanamoothoo
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Umbilical Cord ◽  
Genetic Disease ◽  
Ethical Issues ◽  
Test Tube ◽  
Alternative Methods ◽  
Official Journal ◽  
Human Fertilisation ◽  
Biomedical Field

AbstractSince the advent of test-tube babies, advances in the biomedical field have risen steadily. In parallel, the scientific body has never since ceased to debate the ethical issues that they arise. This is particularly the case regarding saviour-sibling. Saviour-sibling refers to a child who is conceived to cure an older brother or sister suffering from a serious family genetic disease. Therefore, it is meant to give birth to a child who will provide stem-cells taken from the umbilical cord or bone marrow afterwards, to treat an elder sick sibling. In England, this practice has been explicitly allowed by the new Human Fertilisation and Embryology Act 2008 under some strict conditions. In France, this practice, authorized by the Bioethics Law of August 2004 and confirmed by its decree of implementation published in the Official Journal on 23 December 2006, is also strictly regulated. This technique opens up new perspectives and enormous hope. Its legalisation is certainly justified by the suffering of the parents and to avoid that they travel to other States where it is permitted. However, it raises serious psychological ethical and judicial issues. Following an analysis of the English and French laws on saviour siblings, its controversial side will be highlighted, before concluding whether or not this new Pandora’s box which is saviour-sibling, should be closed and other alternative methods encouraged.

Decreased MHC-II Expression and Immune Dysfunction by Mesenchymal Stem Cells in the Bone Marrow of Patients with Myeloproliferative Disorders.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4647-4647
Author(s):  
Marianne D. Castillo ◽  
Pranela Rameshwar
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Hematological Malignancy ◽  
Flow Cytometric Analysis ◽  
Myeloproliferative Disorders ◽  
Bone Marrow Biopsies ◽  
Hematological Disorders ◽  
Mhc Ii ◽  
Immune Mediated

Abstract The etiology of hematological disorders has been studied at the cellular and molecular levels. These studies have led to an understanding of the effects by the bone marrow microenvironment on the pathophysiology of myeloproliferative disorders. The overarching hypothesis states that resident bone marrow Mesenchymal Stem Cells (MSCs) are important in the development of myeloproliferative disorders and are also involved in the development of fibrosis. The specific hypothesis is that MHC-II expression is decreased in patients MSCs. This makes them unable to act as antigen presenting cells and to suppress immune mediated mechanisms that lead to the development of some myeloproliferative disorders. MSCs were expanded from bone marrow aspirates of patients with AML (n=10), CML (n=10), and MDS (n=10). Flow cytometric analysis showed decreased MHC-II expression in MSCs from all patients as compared to MSCs from patients without hematological malignancy. The flow cytometry results were verified in functional studies using the MSCs as stimulators in a one way mixed lymphocyte reaction. Compared to MSCs from non-hematological malignancy patients, MSCs from study subjects showed reduced ability to elicit allogeneic responses. Retrospective analyses of bone marrow biopsies using immunohistochemistry showed an increase in the amount of MSCs in myelofibrosis patients, compared to patients without evidence of fibrosis, alluding to their role in the development of this condition. These results suggest that MSCs may be dysfunctional in patients with hematological disorders. Variations in the immune properties and the increased amount of MSCs in these patients open an avenue for a lingering question on the etiology of the development of some hematological disorders. Do dysfunctions of MSCs precede myeloproliferative disorders and leukemia or does the opposite occur? In summary, we provide insight into the immune-mediated mechanisms related to the pathophysiology of these disorders, which may have clinical implications for future therapies of bone marrow related disease.

Following Autologous Stem Cell Transplantation (1 Yr) Mesenchymal Stem Cells Are Functionally Impaired with Reduced Hematopoietic Support

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 3888-3888
Author(s):  
Catharina Hazenberg ◽  
Fiona A.J. van den Heuvel ◽  
Edo Vellenga ◽  
Annet Z. Brouwers-Vos ◽  
Gerbrig Berger ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Autologous Stem Cell Transplantation ◽  
Population Doubling ◽  
Bone Marrow Biopsies

Abstract Autologous stem cell transplantation (ASCT) is frequently applied in patients with multiple myeloma and malignant lymphoma. Although adequate steady state hematopoiesis with normal peripheral blood counts is attained after ASCT, marked cytopenias may occur in times of stress such as sepsis or re-exposure to chemotherapy. Our group has previously shown impairment of the hematopoietic stem cell (HSC) compartment 1 year post ASCT (pASCT), reflected by reduced HSC frequency and quiescence, and increased ROS production (Haematologica 2013;98:1264). Considering the essential role for mesenchymal stem cells (MSCs) in supporting hematopoiesis, we studied the MSC compartment 1 year post ASCT. Bone marrow biopsies from pASCT patients (n=17) were studied and compared to normal bone marrow from healthy donors (NBM, n=20) by performing immunohistochemistry staining of endothelial cells by CD34 (indicating microvessel density, MVD) and MSCs by nestin, CD146 (Melanoma Cell Adhesion Molecule, MCAM) and CD271 (Nerve Growth Factor Receptor, NGFR). A significant increase in CD271+ MSCs was observed in pASCT bone marrow biopsies compared to NBM (p<0.0001), while the expression of additional markers did not differ between pASCT vs. NBM. MSCs were cultured from the CD34- fraction of bone marrow mononuclear cells, obtained from pASCT patients (n=17) and MSCs derived from NBM (n=20). MSCs were selected by their plastic-adherency and replated to generate MSCs. Although pASCT MSCs and NBM MSCs had similar population doubling times (1.92±0.22 and 3.52±1.02 in passage 4 (P4), pASCT MSCs cultured in vitro demonstrated a change in morphology from the onset of P4. We also observed premature exhaustion of growth in 45% of the studied patients at P5 (vs. 18% in NBM) and increased senescence shown by B-galactosidase staining in P5/P6 (p=0.04). Differentiation assays did not show impairment in differentiation towards osteoblasts or adipocytes of pASCT MSCs. Gene expression analysis on early passage MSCs showed upregulation of pro-inflammatory and cell cycle genes, such as IL6 and p21, in pASCT MSCs compared to NBM MSCs. Co-culture studies with cord blood-derived CD34+ cells on pASCT MSCs showed a significant reduction in output in CFC assays and significant reduction in number of cobblestone-area forming cells in pASCT co-cultures versus NBM (p < 0.05). Given the higher incidence of MDS and AML after ASCT, we questioned whether the observed phenotype of pASCT MSCs resembles MSCs from patients with MDS and AML. Therefore the endothelial and mesenchymal compartments of MDS (n=20) and AML (n=23) patients were studied. An increase in MVD was detected in MDS/AML bone marrow biopsies in contrast to NBM and pASCT (p < 0.05), while the expression of CD146, CD271 and nestin in MDS/AML patients was not significantly increased. 25% of AML MSC cultures showed no growth in the first passage. When MSC growth did occur, the remaining cultures did not show a difference in population doubling time or expansion. However, a change in morphology of MDS/AML MSCs similar to pASCT MSCs was observed. Studies of early passages of MDS/AML MSCs demonstrated a significantly increased gene expression of IL-6 and p21 comparable to pASCT MSCs. In addition PITX2 and Foxc1 expression was increased but no difference was observed in pASCT vs. MDS/AML MSCs. PITX2 has been linked to increased senescence of MDS MSCs while Foxc1 is linked to adipo-osteoprogenitor cell differentiation thereby affecting the HSC compartment. Since none of the pASCT patients did develop MDS, immunohistochemical stainings were also performed on bone marrow biopsies of patients that developed therapy related (t-)MDS/AML following ASCT for lymphoma and myeloma (n=7), after a mean of 117 (MDS) and 50 months (AML). An increase in MVD was observed shortly before or during MDS/AML development, which is probably related to the emergence of malignant cells. No major changes in the phenotype of the MSC compartment were observed before or during the emergence of t-MDS/AML, indicating that t-MDS/AML is preceded by an increase in MVD without distinct changes in the MSC compartment. In summary our results demonstrate that MSCs are affected after ASCT, as shown by expression pattern and functionality. These changes result in a pro-inflammatory phenotype with premature senescence and impaired support of hematopoietic cells, which may account for the reduced bone marrow reserve observed in pASCT patients. Disclosures No relevant conflicts of interest to declare.

scholarly journals Fundamental research for jawbone regenerative medicine development using adipose tissue-derived stem cells

Impact ◽  
2021 ◽  
Vol 2021 (5) ◽  
pp. 34-36
Author(s):  
Goichi Matsumoto
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Adipose Tissue ◽  
Growth Factor ◽  
Maxillofacial Surgery ◽  
Autologous Bone Marrow ◽  
Alternative Methods ◽  
Oral And Maxillofacial Surgery ◽  
Differentiation Potential ◽  
Cell Based Therapy

Bone loss around the teeth and jaw can occur for a number of reasons and this can lead to deterioration of the alveolar and masticatory and aesthetic disorders. Oftentimes, this impairs quality of life. It is possible to reconstruct the lower jaw using bone harvested from the patient's body but there are limitations associated with this method. Another option is using bone marrow cancellous bone fragments containing autologous bone marrow-derived somatic stem cells (BMSCs) but, again, there are limitations. Therefore, alternative methods of jawbone reconstruction are required. Dr Goichi Matsumoto, Department of Oral and Maxillofacial Surgery, Kanazawa Medical University, Japan, is an expert in this area. He is exploring the potential of using adipose tissue-derived stem cells (ADSCs) in jawbone reconstruction and believes this would overcome the limitations of existing methods as well as advancing regeneration therapy. There are numerous benefits to the use of ADSCs, but in order to move forward with this it is first necessary to explore the characteristics of ADSCs in detail, this includes investigating 'the proliferation ability and multi-differentiation potential of ADSCs. This will enable high-quantity and quality ADSCs to be obtained for clinical cell-based therapy and tissue engineering. It is Matsumoto's goal to develop a new mandibular regeneration treatment that involves regenerating the mandible by locally releasing a growth factor with bone-forming ability called basic fibroblast growth factor (bFGF). The researchers are also working to develop and commercialise a hybrid bone cement.

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