bluuebunny/crossref_metadata_2025 huggingface dataset:
Safety, Tolerability, and Pharmacokinetic Assessment of CG53135 Administered Intravenously in Patients with Advanced (Stage 4) Cancer (Single Rising-Dose Tolerance).
Authors: Stephenson Joe , Goel Sanjay , Bhargava Pankaj , Vadhan-Raj Saroj , Annino Vincent , Hawthorne Thomas , Halvorsen Yuan-Di , Hahne William
Published: October 2019

Oral mucositis (OM) is a common complication in cancer patients receiving chemotherapy (CT). OM is characterized by damage to the epithelium of the oral-pharyngeal cavity. CG53135-05, a recombinant human fibroblastic growth factor 20 (rhFGF-20) protein, has demonstrated epithelial and mesenchymal cell proliferation stimulating activity in vitro and reduces OM using single doses in a hamster mucositis model. The goal of this dose-escalating tolerance study was to assess the safety, tolerability and pharmacokinetics of CG53135-05 in cohorts of four patients (pts) at 0.03, 0.1, 0.33, and 1mg/kg (concentrations are determined by the UV method which are equivalent to 0.1, 0.3, 1.0, and 3.0 mg/kg by the Bradford method previously used). Dose escalation was stopped due to tolerability information at 0.33 mg/kg delivered in 15 min (reported in another Phase I study) and the protocol was amended to add a 0.2 mg/kg dose. The World Health Organization (WHO) OM and OMAS scoring systems were used to measure the incidence and severity of OM. The interim results are reported: Ten pts received CG53135-05 at 0.03 mg/kg (n=4) or 0.1 mg/kg (n=6) as a single 100-ml intravenous infusion administered 3 days after completion of the CT. After infusion, CG53135-05 reached maximum plasma concentration within 1h. The mean terminal exponential half-life was 49 min (range: 16.2–87 min, n=5). No pts discontinued the trial due to adverse experiences. Adverse events (number pts) that may be related to the study drug include: nausea (2); chills (2); fever (2); vomiting (1); dizziness (1); vision - “lights flashing” (1) and astigmatism (1); neuropathy (1); tachycardia (1); headache (1); and premature atrial complex (1). All reported incidences were mild to moderate. No grade 3 or 4 laboratory toxicity associated with the study drug was noted. Three serious adverse events deemed unrelated to study drug were noted from two pts including cancer progression, catheter infection, and small intestinal obstruction. Among the treated pts, no grade 3 or 4 OM was observed. Three pts developed Grade 2 mucositis that lasted between 1–5 days. No pts required total parenteral nutrition. Approximately 72–84% of pts receiving CPT-11 experience diarrhea. Of the six pts who received CPT-11 as part of CT in this trial, two pts (both receiving 0.03mg/kg of CG53135-05) experienced mild to moderate diarrhea. Conclusions: CG53135-05 is well tolerated with single dose administration at 0.03 and 0.1 mg/kg. Dosing at 0.2 mg/kg is ongoing. The full results of this study will be presented.
The Suppressive Effect of Bone Marrow Mesenchymal Stem Cells on T Cell Activation, Is Deficient in Patients with Severe Aplastic Anemia.
Authors: Bacigalupo Andrea , Valle Marisa , Podesta’ Marina , Pitto Anna , Zocchi Elena , De Flora Antonio , Pozzi Sarah , Vassallo Franca , Lucchetti Silvia , Figari Osvaldo , Frassoni Francesco , Piaggio Giovanna
Published: October 2019

Normal bone marrow derived mesenchymal stem cells (MSC) suppress T cell function, and may provide an immunoprotected environment for hemopoietic stem cells. In this study we compare the suppressive activity on T cell function of MSC from 23 patients with severe aplastic anemia (SAA) patients, at diagnosis (n=3), following immunosuppressive therapy (IS) (n=16) or after an allogeneic bone marrow transplant (BMT) (n=4), and normal individuals. As shown in Figure 1 increasing numbers of control MSC (25, 50 and 100x10^3 cells) produced a dose dependent suppression of PHA induced T cell proliferation; when MSC were derived from SAA patients, they exhibited significantly less suppressive activity (37% vs 6%, 81% vs 24%, 96% vs 35%). The reduced ability of SAA MSC to suppress mitogen induced T cell proliferation, was seen irrespective of disease phase. Paired experiments on mixed lymphocyte reaction confirmed the inability of SAA MSC to suppress T cell function. Other abnormalities of SAA MSC included: (a) impaired capacity to down regulate CD38 expression on PHA primed T cells, (b) impaired ability to suppress gamma-IFN production in PHA coltures, which resulted in a 11 fold different gamma-IFN concentration in the cultures; (c) no effect on T cell mediated inhibition of hemopietic colony formation. Finally SAA MSC produced significantly (1 log) less adenosin diphosphate -ribosyl cyclase (cADPR), a promoter of in vitro hematopoiesis. MSC mediated suppression of PHA induced T cell proliferation, was restored to control levels in 3 of 4 patients post-BMT. In conclusion, the ability of MSC to down regulate T cell priming, proliferation and cytokine release is deficient in patients with SAA, persists indefinitively after immunosuppressive therapy, but may be restored after BMT. Whether this defect plays a role in the pathogenesis of marrow failure, remains to be determined.

Figure Figure
Results of a Phase I/II Randomized Immunotherapy Trial Testing Adoptive Transfer of Co-Stimulated Autologous T Cells and Pneumococcal Conjugate Vaccine (PCV) Immunization after Autotransplantation for Myeloma (MM).
Authors: Cross Alan , Rapoport Aaron P. , Levine Bruce L. , Stadtmauer Edward A. , June Carl H. , Mann Dean , Frasch Carl , Lynch Christian , Badros Ashraf , Chrisley Lisa , Cotte Julio , Mair Rebecca , Hinkle Joanne , Zhang Lei , Edelman Robert
Published: October 2019

Although immunizations against microbial infections are recommended following autologous stem cell transplantation (ASCT), the antibody responses are often suboptimal. To assess improved immunization strategies, we tested the ability of autologous T cells that were co-stimulated ex vivo with anti-CD3 and -CD28 coated beads as part of their therapy for MM, as well as pre-ASCT immunization to enhance the antibody response to PCV post-ASCT. Subjects were randomized to receive PCV (groups 1 and 2) or no immunization (groups 3 and 4) before T cell harvest, and subsequent mobilization chemotherapy. All subjects received PCV at days 30 and 90 post-ASCT; however, groups 1 and 3 were given T cells before (day 12) post-ASCT PCV immunization, while groups 2 and 4 received T cells after PCV immunization (day 100). Patients receiving adoptive T cell therapy for MM without PCV immunization (group 5) served as controls. The results of the adoptive transfer of T cells on MM therapy are reported elsewhere. Anti-pneumococcal (Pn) antibody levels for serotypes 6B, 14, 23F, 19F were measured in sequential samples for up to 180 days for each patient by ELISA (response defined as ≥ 2-fold increase over baseline and ≥ 0.5 μg/ml) and opsonophagocytosis assay (OPA) (response defined as ≥ 1-log bacterial kill or ≥ 70% kill when ≤ 30% kill at baseline).

Groups 1 2 3 4 5 a Numbers of evaluable subjects per group responding by ELISA to 3 or more of 4 pneumococcal serotypes tested. bNumber of evaluable subjects with OPA response to 19F only. T=T cells; D=day;* P=0.02, and** P=0.05 v. group 5 by Fisher’s Test. No. PCV 3 3 2 2 0 T given at D12 D100 D12 D100 D12 ≥3 types 6/7a 3/8 3/9 3/6 1/4 4 types 6/7* 1/8 1/9 3/6 0/4 OPA 8/9b** 5/8 1/8 3/5 1/4

Responses in group 1 were of longer duration and greater magnitude than in other groups. We conclude that (1) MM patients immunized with PCV pre-ASCT respond better to PCV post-ASCT; (2) T cell administration before PCV immunization enhances protective anti-Pn antibody levels and for a longer duration. Our vaccine schedule that leads to improved PCV responses may also be used to improve the responses of ASCT patients to other microbial and perhaps cancer vaccines. (This study was supported by NIH contract N01-AI-85342 and LSA SCOR #7000-002).
Control of Fulminant Juvenile Myelomonocytic Leukemia (JMML) Relapse and Gut GvHD III° with Purinethol Early after Unrelated Donor BMT.
Authors: Stachel Daniel , Gravou-Apostolatou Chara , Kuhlen Michaela , Leipold Alfred , Bader Peter , Beck Jorn , Holter Wolfgang
Published: October 2019

Juvenile myelomonocytic leukemia (JMML) is a rare disease in childhood which can only be cured by stem cell transplantation. The major complication is relapse in up to half of the patients. The existance and efficacy of graft-versus-leukemia (GvL) in JMML is controversial and often associated with severe graft-versus-host disease (GvHD).

A 1,5 year old boy developed JMML and was transplanted from a 1 antigen mismatched UD (unmanipulated bone marrow, 8.5 Mio CD34/kg) after a CR consisting of Bu 16*1.25 mg/kg), Cy (2*60 mg/kg), Mel (1*140 mg/m2) and ATG (3*20 mg/kg). GvH prophylaxis consisted of CsA and very short MTX. The situation was further complicated by the intermittent presence of CMV, HHV-6 and EBV in the peripheral blood which was treated intermittently by intravenous ganciclovir.

Engraftment occurred on day + 16. GvHD III° of the skin only developed and was treated with corticosteroids, CsA and MMF. Chimerism was complete on day +28.

Beginning on day +45 an increasing autologous chimerism was detected. Therefore, immunosuppression was halted. Despite discontinuation of all immunosuppressants the autologous chimerism increased to 60–80% (d +63) and the peripheral leukocytes increased to approx 30,000/μl together with eosinophilia (d +60). Clinical signs of relapse (hepatomegaly and pulmonary obstruction) were also present. Thereafter, within a week, leukopenia and thrombocytopenia developed and the autologous chimerism decreased to 1–5%.

Coinciding with the apparent GvL effect severe GvHD reappeared. Skin GvHD II–III° developed, than gut GvDH III° with massive life threatening fluid and potassium loss (day +73).

In an attempt to treat both JMML and GvHD the antimetabolite purinethol 50 mg/m2 daily was given orally.

Since day + 98 always an complete chimerism was observed. Gut GvHD gradually improved without further immunosuppression. The boy is now at home without evidence of disease or active GvHD more than 1 year after relapse.

We speculate that in this case purinethol controlled not only the severe gut GvHD after BMT but also JMML.

This antimetabolite may therefore be considered as an immunosuppressant for GvHD when malignat relapse is also present or imminent.
Blood Stream Infection after Reduced- Intensity Unrelated Cord Blood Transplantation for Adults with Hematologic Diseases.
Authors: Narimatsu Hiroto , Kami Masahiro , Matsumura Tomoko , Myojo Tomohiro , Masuoka Kazuhiro , Inokuchi Chiho , Yuji Koichiro , Miura Yuji , Kusumi Eiji , Ueyama Junichi , Wake Atsushi , Miyakoshi Shigesaburo , Yoneyama Akiko , Matsuzaki Michio , Morinaga Shinichi , Muto Yoshitomo , Taniguchi Shuichi
Published: October 2019

Blood stream infection (BSI) is a major problem after cord blood transplantation (CBT). However we have little information on it after reduced-intensity cord blood transplantation (RICBT).

This study aims to investigate clinical characteristics of BSI within 100 days of RICBT, and to identify its risk factors in the patient who received RICBT between January 2002 and March 2004 in Toranomon hospital.

We reviewed medical records of 77 patients with advanced hematologic diseases who underwent RICBT at Toranomon Hospital between January 2002 and June 2004.

Median age of the patients was 55 years (17-79). Median number of infused cells and CD34+ cells were 0.86 x 10E7 (range; 1.73–4.31) and 0.8x10E7/kg (range; 0.01–46.1), respectively. Conditioning regimen consisted of fludarabine (125–150 mg/m2), melphalan (80–140 mg/m2), and TBI 2–8 Gy. GVHD prophylaxis was cyclosporine or tacrolimus. BSI must have met at least one of the following criteria previously described. (O’Grady, NP et al., Infect Control Hosp Epidemiol 2002)

Mortality was considered to be directly attributable to a bloodstream pathogen if the patient died within 7 day after the last positive blood culture without any other probable cause of death, The patient’s characteristics were compared between those with or without BSI by univariate analysis.

149 episodes of BSI were observed in 31 patients. Cumulative incidence within 100 days was 0.40. Median onset was day 13 (1–98) after RICBT.

The causative organisms were P. aeruginosa (47), S. epidermidis (44), E. faecium (13), T. beigelii (6), S. maltophilia (6), E. faecalis (5), E. coli (3), A. xylosoxidans (3) and others (22). 8 patients died of septicemia directly associated with BSI. A mortality rate was 26%. The causative organism included P. aeruginosa, A. xylosoxidans, S. maltophilia, S, aureus and gram-positive rod. Use of corticosteroid (p=0.012) and time to engraftment (p=0.003) were associated with the development of BSI.

BSI is a major complication in our series of RICBT. Use of corticosteroid and prolonged neutropenia might have increased the risk of incidence of BSI.
The Zebrafish Hypochromic Mutant [iItalic]Shiraz[/iItalic] Encodes a Novel Mitochondrial Glutaredoxin That Establishes a Link between Heme and Fe/S Production.
Authors: Wingert Rebecca A. , Barut Bruce , Foott Helen , Fraenkel Paula , Dooley Kimberly , Kingsley Paul , Palis Jim , Shubert Heidi , Chen Opal , Kaplan Jerry ,   Tubingen 2000 Screen Consortium, Zon Leonard I.
Published: October 2019

Iron is required in the mitochondria both to produce heme, which is used for hemoglobin synthesis, and to make iron-sulfur (Fe/S) clusters, which confer electron transfer or catalytic functions to proteins. Cellular iron utilization and Fe/S cluster production are thought to occur independently, yet the processes are coordinated through currently uncharacterized pathways. The shiraz (sir) zebrafish mutant manifests a hypochromic, microcytic anemia. Positional cloning of sir discovered a deletion at the locus that included the zebrafish orthologue to glutaredoxin 5 (grx5), a gene required in yeast for Fe/S cluster assembly. We found that grx5 is highly expressed in the developing blood and fetal liver of both zebrafish and mouse embryos. Antisense-mediated morpholino knockdown of grx5 prevented hemoglobin production, and overexpression of zebrafish, yeast, mouse, or human grx5 RNA in sir embryos completely rescued hemoglobin production, indicating that grx5 is the gene responsible for the sir phenotype. Expression of zebrafish grx5 was found to rescue Fe/S protein production in the yeast Δgrx5 strain, demonstrating that the role of grx5 in Fe/S cluster assembly is conserved among eukaryotes. The surprising finding that mutating a gene necessary for Fe/S cluster assembly caused a lack of hemoglobin synthesis suggested that we had discovered a connection between these pathways. In vertebrates, iron regulatory protein 1 (IRP1) acts as a sensor of intracellular iron levels and controls cellular iron homeostasis via posttranscriptional regulation of iron uptake, storage, and utilization genes. For instance, IRP1 binds to the 5′ iron response element (IRE) in the aminolevulinate synthase 2 (ALAS2) mRNA, blocking translation when cellular iron is low. However, when cellular iron is replete, IRP1 binds a Fe/S cluster and its RNA-binding activity is abolished. We hypothesized that the loss of Fe/S cluster assembly in sir would activate IRP1 and block ALAS2 synthesis, resulting in hypochromia. In support of this model, overexpression of ALAS2 RNA without the 5′ IRE rescued sir hypochromia, while overexpression of ALAS2 including the IRE did not facilitate rescue. Furthermore, antisense morpholino knockdowns of IRP1 caused rescue of hemoglobin synthesis in sir embryos. The combination of these data indicate that hemoglobin production in the differentiating red cell is monitored through Fe-S cluster assembly as a mechanism to gauge iron levels and accordingly direct heme synthesis. This finding illustrates a crucial role for the mitochondrial Fe/S cluster assembly machinery during hemoglobin production, and has broad implications for the role of such genes in human hypochromic anemias.
Haploidentical Hematopoietic Cell Transplantation (HCT) Compared with Matched HCT from Family Donors: report of 216 cases.
Authors: Lu Daopei , Han Wei , Dong Lujia , Huang Xiaojun , Liu Kaiyan , Chen Huan , Xu Lanping , Ren Hanyun , Liu Daihong , Chen Yuhong , Wu Tong , Lu Jin , Gao Zhiyong , Wang Jingbo , Jiang Qian , Zhang Yaochen , Wang Jingzhi
Published: October 2019

A total of 216 cases of matched and mismatched-haploidentical HCT from family donors have been performed since May 2002 in our Institute. The purpose of this analysis is to compare the GvHD, relapse rate and their risk factors for complications and survival. The feasibility of the present regimen can then be evaluated. In the arm of mismatched-haploidentical HCT, GIAC regimen (G-CSF priming hematopoietic cells collection; immunosuppression intensified and prolonged; ATG being used; combination use of BM + PB) was used for the first time. It was developed for patients without HLA matched related or unrelated donors. However, in HLA matched HCT, ATG was not used. The two groups were comparable in disease diagnoses, sex, and prophylaxis of GvHD, number of MNC/kg and use of G-CSF post-transplant. The great majority of recruited patients had hematological malignancies. A few were cases of SAA. There were significantly more patients in advanced stage or in high-risk status in mismatched-haploidentical HCT group. After median value of 9(2–260 months follow up, the results are shown in Table 1.

Table 1. Survival and causes of Death (2-year Kaplan-Meire Estimates)

Characteristics and Outcomes Matched Mismatched-haploidentical No. Of Patients 116 100 Age (yr.) 37 (12–62) 23 (3–52) Status of Patients Standard Risk 86 (74.8%) 44 (44%) High Risk 30 (25.2%) 56 (56%) Days post-transplant ANC>0.5x109/L 16.4 12 Platelets>20x109/L 16.9 17 Acute GvHD <100 days 0-I 48.6% 52% II 38.6% 35% III-IV 12.8% 13% Chronic GvHD 62.5% 63.3% Extensive 18.7% 18.3% Overall survival for 1 year 81.2% 72% Relapse related mortality 5.17% 6% Non-relapse related mortality 11.2% 16%

In summary, compared to matched HCT, GIAC regimen for mismatche-haploidentical HCT is sufficiently safe for patients.
Comparison of Outcomes of Unrelated Bone Marrow and Umbilical Cord Blood Transplants in Children with Hematologic Malignancies: Single Center Study in Korea.
Authors: Chung Nak-Gyun , Cho Bin , Park Young-Shil , Jeong Dae-Chul , Jang Pil-Sang , Kim Hack-Ki
Published: October 2019

In order to compare the outcomes of unrelated umbilical cord blood transplants (UCBT) or bone marrow transplants, 102 children with hematologic malignancies transplanted with unrelated umbilical cord blood (n=35, M:F=21:14, median age 4 years) or unrelated bone marrow transplants (UBMT) (n= 67, M:F=49:18, median age 9 years) were analyzed in a retrospective single center study (between Aug. 1997 and Dec. 2003 in the Catholic Hematopoietic Stem Cell Transplantation Center of Korea). HLA mismatches were defined by serology for class I and molecular typing for DRB1. The donor was HLA mismatched in 94% of UCBTs (33 of 35 UCBT) and in 0 % of UBMT. Non-adjusted estimates of 2-year event-free survival rates were 46 % and 57 %, respectively (P > 0.2). Engraftment failures of donor cells occurred in 17 % (6 out of 35) of UCBT and in 6 % (4 out of 67) of UBMT. Grade >= III acute GVHDs developed in 3 out of 29 (10 %) engrafted patients who underwent UCBT and in 17 out of 63 (27 %) engrafted patients who underwent UBMT. Early TRM at day 100 was 31 % (11 out of 35) for UCBT and 22 % (12 out of 67) for UBMTs. Within post-transplant month 3, CMV infections occurred in 37 % of UCBT and 54 % of UBMT, but most of them did not progress to CMV disease due to early ganciclovir treatment. Chronic GVHD developed in 2 out of 22 (9 %) patients who underwent UCBT and in 15 out of 49 (31 %) patients underwent UBMT.

In conclusion, the use of UCBT as a source of hematopoietic stem cells is a reasonable option for children with hematologic malignancies lacking an acceptable, matched unrelated marrow donor.
Myeloablative Conditioning in Myelofibrosis using i.v. Treosulfan and Autologous Peripheral Blood Progenitor Cell Transplantation with High Doses of CD34+ Cells Results in Hematologic Responses - Follow-Up of Three Patients.
Authors: Fruehauf Stefan , Buss Eike C. , Topaly Julian , Kreipe Hans H. , Ho Anthony D.
Published: October 2019

Myelofibrosis with myeloid metaplasia is a chronic myeloproliferative disorder. The only curative therapy to date is myeloablation followed by allogeneic transplantation. This approach, however, is associated with a high morbidity and mortality. Autologous transplantation after myeloablation with high-dose oral busulfan provides a palliative approach which can lead to a long-term relief of symptoms and is associated with acceptable morbidity and mortality (Anderson JE et al. 2001, Blood 98: 586). However, busulfan pharmacokinetics after oral administration can vary between patients and increased toxicity is encountered in some. On the other hand treosulfan, a bi-functional alkylating drug, can be administered safely i.v. with reliable pharmacokinetics up to the myeloablative dose range. In the study presented here, patients were stimulated with G-CSF 16 μg/kg daily for 4 days and subsequent leukapheresis of a minimum of 5 x 106 CD34+ cells/kg was performed. Myeloablation consisted of treosulfan infusions of 14 g/m2 for three consecutive days (total dose 42 g/m2) and subsequent autologous PBPCT. To date we have transplanted 3 patients, all female. Two patients (#1, #3) had symptomatic splenomegaly and severe anemia. Patient #2 had symptomatic splenomegaly and thrombocytopenia (< 100/nl). The WB nadir after treosulfan therapy was 0.06/nl (#1), 0.28/nl (#2) and 0.09/nl (#3). The time to reconstitution of leucocytes > 1/nl post transplantation was 28 days (#1, #2) and 38 days (#3) and the time for reconstitution of thrombocytes > 50/nl was 36 (#1), 22 (#2) and 33 (#3) days. The prolonged reconstitution period may have been due to the myelofibrosis and has also been observed after busulfan conditioning and PBPCT by others. There were no fever or other severe toxicity. Patients did not require total parenteral nutrition. The patients have been followed-up for 18 months post transplantation, now. The first patient (#1), who required erythrocyte transfusions twice weekly pretransplant received her last erythrocyte transfusion on day 56; her Hb-value is 11.2 g/dl at last follow-up. The second patient (#2) recovered to platelet counts higher than pre-transplantation (58/nl) at 3 months.(143/nl) and had 103/nl at last follow-up. Pat. #3 showed a marked reduction of max. spleen size and a rise in Hb from 9 g/dl to 12 g/dl after 12 months, the Hb value at 18 months is 8.7 g/dl and allogeneic transplantation is considered now. We conclude that myeloablation with treosulfan and autologous PBPCT in these three patients with myelofibrosis was safe and useful. Survival of these patients is promising up to date. The results presented warrant further exploration of this approach in a clinical study which has been initiated by our group. (cf. http://leukaemie.krebsinfo.de/kn_home/Studien/studie_101.html).
Stem Cell Transplantation in 281 Patients over 60 Years in Onco Hematologic Malignancies: A Single Center Experience.
Authors: Stoppa Anne-Marie , Vey N. , Faucher C. , Bouabdallah R. , Viret F. , Chabannon C. , Ladaique P. , Coso D. , Charbonnier A. , Schiano J.M. , Viens P. , Gastaut J.A. , Maraninchi D. , Blaise D.
Published: October 2019

From 06/1982 to 12/2003, 281 patients (163 male) received autologous (AuSCt) and/or allogeneic (AlloSCT) transplantations as part of first line treatment (61%) second line treatment(21%) or salvage therapy (17%). There were 140 multiple myelomas (including 3 amyloidosis), 62 lymphomas, 51 acute myeloid leukemias, 24 solid tumors and 4 chronic myeloid leukemias.Two hundred and sixty nine patients-median age: 64 y(60–77) - received Au SCT (28/261 received a second Au SCT in med of 2–7 months), 1patient (71 y) received a double syngeneic transplant,12 patients -median age: 62(60–68) received AlloSCT. Conditionning regimen (Cond reg) for the first Au SCT included Melphalan 140 mg/m² n=168; BEAM n=41; Melphalan 200mg/m² n=29; Mel Busulfan (8 to 16 mg/kg) or Mel TBI 8 grays n=18; Cytoxan 200mg/kg Melphalan1 40 n=9: TBI 8 Grays n=7; BCNU 600mg/m² n=3. Cond reg for the second Au SCT included Mel 200 n=20; Mel 140 n=4; BCU 600 n=4. Cond reg for the double syngeneic transplant were Mel 140 and Bu 8 Mel 140.Cond reg for the AlloSCT were Fludarabin/Busulfan/ATG (FBS) based reduced intensity regimen (RIC) in 11 cases. Ninety percent of the patients received peripheral blood stem cells-med 5,3.10*6CD34/kg (2–40)- 10% of the patients transplanted before 1994 received bone marrow cells. Tolerance was good with only 9/281 transplant related deaths: 5 sepsis/MOF in aplasia (D11–D21) 2 ARDS (D30–D80) 1 AGVH (D 60) 1 CGVH (3 years). Theses deaths occured in 4 ref MM( 2 renal instability,1 cardiac amyloidosis), 2 CR2 NHL, 1 OR1 MM, 1 stable MDS after FBS in 2 pts, BEAM in 2 pts, Mel 140 in 4 pts, Mel 200 (second transplant) in 1 pt. With a median follow up of 20 months from transplant,7/281 second cancers had occured: 1AML, 1ALL, 2 NSC pulmonary carcinoma, 1 colon cancer, 1uterin carcinoma, 1 melanoma; 2 of these 7 patients had died. One hundred and fifty eight patients had relapsed in a median of 11 months (1–92). Median survival for the 281 patients is 37 months without diference between pts older or younger than 65. Median survival for MM, NHL/HL, AML or ST are 40 months, 45 months, 18 months and 23 months respectively
Targeted Radiotherapy to the Skeleton Using 166Ho-DOTMP with Autologous Stem Cell Transplantation for Patients with Bone-Only Metastatic Breast Cancer.
Authors: Champlin Richard , Booser Daniel , Rondon Gabriela , Williams Patricia , Wendt Richard , Hortobagyi Gabriel , Ueno Naoto , Podoloff Donald
Published: October 2019

The skeleton is the most frequent site of metastatic disease in breast cancer, involving cortical and trabecular bone with or without extension into adjacent bone marrow. A subset of breast cancer patients develop bone dominant disease without metastases to other organs over a prolonged period of time. An attractive strategy is to utilize bone targeted radiotherapy to address this site of disease with minimal systemic toxicity. This study was designed to determine the safety and preliminary efficacy of 166Ho-DOTMP, which localizes to bone providing radiation to adjacent marrow and malignant cells. The limiting toxicity is myelosuppression, which can be overcome by autologous stem cell transplantation. We studied this strategy in subjects with breast cancer metastatic to confined to the bone.

Treatment occurred between 1997 and 1999. Subjects were eligible if they were <65 years of age, had bone only disease, had an estrogen receptor negative tumor or had failed hormonal therapy. A 30 mCi tracer dose was administered to assess targeting and to allow the therapeutic dose to be calculated.

If uptake was adequate, a therapeutic dose was to be administered in one of 2 dose levels, receiving 22 Gy (n=3) or 28 Gy (n=3). Treatment was followed by autologous stem cell transplantation when the ongoing radiation dose to marrow was <1 cGy/hour. The 6 subjects received a median of 1548 mCi (range 870 2065 mCi) of 166Ho DOTMP. There was selective uptake in the bone, particularly in sites of osteoblastic metastases, with unbound drug cleared into the urine. The most common toxicities seen during active treatment were mild nausea and vomiting. None of the patients experienced grade >3 acute toxicity aside from the expected profound myelosuppression. All subjects treated in the trial had prompt trilineage hematologic recovery post transplant. Two patients developed hemorrhagic cystitis at approximately two years following therapy which resolved in both patients. One of these patients also had GI bleeding and pseudomembranous colitis unrelated to the 166Ho DOTMP treatment. One subject developed MDS, but was found to have a pre-existing trisomy 8 by FISH analysis from the peripheral blood stem cell product collected prior to treatment on this study. Two patients achieved complete remission and remain progression-free over 5 years following study entry. Four relapsed and died of progressive disease; all recurred in extraosseous sites with two also having disease in the bone marrow. Median time to progression was 10 months.

Conclusions: Treatment with 166Ho DOTMP delivers targeted radiotherapy to the skeleton with an acceptable toxicity profile. Two of 6 patients with bone-only metastases achieved sustained complete responses. Further studies are warranted to evaluate this strategy for treatment of bone metastases.
Dendritic Cells Tranfected with CML-28 cDNA Induce CML28-Specific Cytotoxic T Cells.
Authors: Zhou Hongsheng , Zhang Donghua , Wang Yaya , Xiao Yi , Huang Wei , Li Dengju
Published: October 2019

Dendritic cells (DC) are the most potent antigen-presenting cells known; the use of DC loaded with tumor antigen is one of the most promising approaches to induce specific immune response. CML28 is an attractive candidate. According to the published article and our following study, CML28 is a broadly immunogenic antigen, which is overexpessed in leukemia cells, including acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL) and chronic myelogenous leukemia (CML); but not in normal hematopoietic tissues or other normal tissue, with the exception of testis. In our study, DCs were generated from peripheral blood mononuclear cells (PBMC) by culture with GM-CSF, IL-4 and TNF-α. The full CML28 cDNA was amplified from K562 cell and then cloned into plasmid pcDNA3.1 HisA. Load DC with the constructed plasmid pcDNA3.1 HisA-CML28 by electroporation. The results of FACS analysis and Western Blot respectively demonstrate that the plasmid transfected DC retain immunophenotypical characteristics and can express the fusion protein CML28-His. Cytotoxic T lymphocytes (CTL) were generated from autologous PBMC stimulated by the transfected DCs. Cytoxicity assay in vitro revealed that CTL can recognize and lyse CML28-expressing target cells, such as the transfected DCs but not normal autologous DCs, which show that the plasmid pcDNA 3.1 HisA-CML28 transfected DC can induce CML28-specific CTL. Our studies demonstrate that CML28 can be a promising target of tumor antigen for leukemia-specific immunotherapy and DC loaded with CML28 tumor antigen can be a promising tool to induce specific anti-leukemia immune response.
MLL Rearranged Infant Acute Lymphoblastic Leukemia Is Characterized by Silencing of the Putative Tumor Suppressor Gene FHIT.
Authors: Stam Ronald W. , Den Boer Monique L. , Passier Monique , Sallan Stephen E. , Armstrong Scott A. , Pieters Pieters
Published: October 2019

MLL gene rearranged acute lymphoblastic leukemia (MLL) is an aggressive type of acute lymphoblastic leukemia (ALL) with a relatively poor response to treatment, predominantly occurring in infants (<1 years of age). Since conventional chemotherapy is failing in >50% of these patients, identification of novel therapeutic targets to direct therapy against is demanded. For this, understanding this disease and finding biological and molecular features characterizing MLL may be crucial. Here we report that infant MLL is characterized by silencing of the putative tumor suppressor fragile histidine triad (FHIT) gene.

Gene expression profiling showed that FHIT expression is significantly reduced in MLL patients (n=20) as compared to patients diagnosed with conventional ALL (n=24) bearing germ line MLL genes. Using quantitative real-time PCR we confirmed FHIT mRNA expression to be low in MLL gene rearranged infant ALL (infant MLL) as compared to both infants and older children (non-infants) not carrying chromosomal abnormalities involving the MLL gene. Infant MLL patients (n=35) significantly (p<0.001) expressed ~15-fold less FHIT mRNA compared to infant ALL patients (n=8) and 11-fold less than non-infant ALL patients (n=22).

Methylation specific PCR (MSP) analysis demonstrated that the FHIT promoter region is hypermethylated in all of the 36 infant MLL cases tested, resulting in strongly reduced FHIT mRNA as well as FHIT protein expression. In contrast, the incidence of FHIT promoter methylation in both infant and non-infant ALL patients carrying germ line MLL genes was markedly lower, with a frequency of 63% (5/8) and 56% (24/43) respectively. Suppression of FHIT expression could be reversed by exposing leukemia cells bearing MLL gene rearrangements to the demethylating drug 5′-aza-2′-deoxycytidine.

These findings imply that MLL rearranged ALL is characterized by FHIT promoter hypermethylation. Currently experiments are being conducted to determine the biological significance of FHIT silencing in this type of cancer.
An Novel Approach in Protective Gene Therapy: Recombinant Adeno-Associated Virus 2-Mediated Transfer of the Human Superoxide-Dismutase Gene.
Authors: Veldwijk Marlon R. , Herskind Carsten , Laufs Stephanie , Stiefelhagen Marius , Zeller W. Jens , Fruehauf Stefan , Wenz Frederik
Published: October 2019

Background and purpose: The success rate of any therapeutic approach depends on the therapeutic window, which can be increased by either raising the resistance of the normal tissue such as hematopoietic cells without protecting the tumor cells or by sensitizing the tumor cells but not the normal cells. Two promising candidate genes for normal tissue protection against superoxide-induced damage may be the copper-zinc (CuZnSOD) and manganese superoxide-dismutase genes (MnSOD). Oxidative stress-induced apoptosis plays a role in both radiation- and chemotherapy-induced tissue damage. Relevant cytostatic drugs for which oxidative stress as one mechanism of action has been described are e.g. anthracyclins, platinum analogues, etoposide and ara-C. For gene therapeutic approaches, recombinant adeno-associated virus 2 (rAAV-2)-based vectors offer attractive advantages over other vector systems: low immunogenicity, possibility of in vivo application, ability to infect dividing and non-dividing tissues and a low chance of insertional mutagenesis, due to extra-chromosomal localization. Here, we report the production and testing of novel rAAV-2-SOD vectors with the goal of normal tissue protection.

Material and methods: Various rAAV-2 vectors containing CuZnSOD, MnSOD and fusion proteins of both with the enhanced green fluorescent protein (eGFP) gene were cloned and vector stocks were produced. Human cervix carcinoma (HeLa-RC) cells were chosen for their susceptibility to rAAV-2. Cells were seeded and transduced with the rAAV-2-SOD vectors. Gene transfer and transgene expression were investigated using FACS and an SOD-activity assay.

Results: Over 70% of all HeLa cells expressed SOD and significant amounts of functional SOD protein were detected (table 1).

Conclusion: These results forms the basis to evaluate the radio- and chemoprotective effects of AAV-mediated SOD gene therapy in hematopoietic and non-hematopoietic (e.g. mucosal) cells.

Vector % GFP+ cells SOD activity (U/mg) Mock control - 435 ± 37 rAAV-2-CuZnSOD N/A 965 ± 112 rAAV-2-CuZnSOD/eGFP 78 ± 2 1093 ± 178 rAAV-2-MnSOD N/A 1516 ± 191 rAAV-2-MnSOD/eGFP 77 ± 3 1204 ± 124
Simultaneous Silencing of Two Independent Signaling Pathways Essential for Angiogenesis Using Bispecific, Tetravalent Intra-Diabodies.
Authors: Jendreyko Nina , Popkov Mikhail , Rader Christoph , Barbas Carlos F. , Gaedicke Gerhard
Published: October 2019

Vascular endothelial growth factor and its receptors (VEGF-R) have been implicated in promoting tumor growth and metastasis. Emerging evidence suggest that angiogenesis also play an important role in the growth of leukemias and lymphomas. Antiangiogenic therapies that interfere with the VEGF/VEGF-R pathways represent novel approaches to effective treatment for certain leukemia, such as myeloid leukemias. But not all types of tumors respond to interruption of the VEGF/VEGF-R pathway. This suggest that alternative pathways for vascular growth exist and can drive tumor angiogenesis. Intrabodies have been used to specifically target intracellular proteins and manipulate biological processes,as well as for the generation of phenotypic knockouts in vivo by surface depletion of extracellular or transmembrane proteins. In order to generate a precise tool for the simultaneous silencing ot two independent signaling pathways essential for angiogenesis, we developed an endoplasmatic-reticulum (ER) targeted bispecific, tetravalent intrabody format, targeting VEGF-R2 and Tie-2. Comparison of the ER-targeted intradiabody with the corresponding conventional ER-targeted single-chain antibody fragment (scFv) intrabodies showed that the intradiabody is significantly more efficient with respect to efficiency and duration of surface depletion of VEGF-R2 and Tie-2. In vitro and in vivo data demonstrate the development of a very effective antiangiogenic intrabody format for the simultaneous functional knockout of two cell surface receptors using an adenovirus-mediated gene delivery system. Our findings suggest that simultaneous interference with the VEGF and the Tie-2 receptor pathway results in at least additive antiangiogenic effects. Our study provides a novel antiangiogenic therapeutic tool for the silencing of two independent signaling pathways involved in angiogenesis.
Bone Marrow Biopsy Needle Design Influences the Bone Content of Recovered Specimens.
Authors: Goldenberg Alec , Kelley Priscilla , Liu Cynthia , Sen Filiz , Ibrahim Sherif
Published: October 2019

Introduction: The integrity and composition of bone marrow core biopsies may be influenced by many parameters including patient age, sex, bone density, operator experience, needle selection and biopsy technique. Biopsy needle design impacts on a needle’s performance and the quality of recovered specimens. We studied the effect of modifying the Snarecoil bone marrow biopsy needle tip on the recovery of trabecular bone in core specimens.

Methods: 7 patients, ages 35–92, M/F = 4/3, (2 leukemias, 1 lymphoma, 1 cytopenia, 1 anemia, 2 other) underwent a bone marrow biopsy using a 8 gauge Snarecoil needle with a new needle tip (TPNSP). The tip has a twin peak point and more distal Snarecoil position. As a control group, 7 patients biopsied with a standard 8 gauge Snarecoil needle (NTP) were retrospectively identified and matched for age and sex and were ages 37–87, M/F = 5/2, (3 cytopenias, 2 leukemias, 1 lymphoma, 1 other). All biopsies were completed using a minimally manipulative technique from the left posterior superior iliac spine with 2% lidocaine. The length of the specimen and attached clot was measured. The specimens were decalcified for 90 minutes and otherwise processed in the standard fashion. The percentage of bone per core biopsy was determined with computer-assisted morphometry using a BioGenex iVision V3.5 system.

Results: The study and control groups were comparable, as the mean age and M/F distribution of the two groups and the mean lengths of the recovered specimens were not statistically significantly different. (TPNSP vs. NTP, 63.8±7.1 vs. 64.7± 6.4 p=0.93, 4/3 vs.5/2 p= 0.57, 2.32± 0.26cm vs. 2.21± 0.26cm p=0.761, respectively).

Needle Age M/F Specimen Length % Bone TPNSP 63.8±7.1 4/3 2.32±0.26 (1.2–3.3) 16.3±1.5 (12.1–21.8) NTP 64.7±6.4 5/2 2.21±0.26 (1.3–2.9) 6.8±1.4 (2.9–12.5)

However, the mean percentage of bone in the specimens recovered by the new TPNSP needle was greater than that recovered with the NTP needle. (16.3±1.5% vs. 6.8±1.4%, p= 0.0008). Grossly, the TPNSP specimens had a more linear edge than those of the NTP specimens. Also, more (5/7) of the NTP specimens had > 0.5 cm of attached clot then did the TPNSP specimens (0/7), p=0.005 suggesting that the twin peak needle produced less tissue hemorrhage.

Conclusions: 1. 8 gauge Snarecoil needles having a sharper tip and more distal Snarecoil recover specimens with as much as 2.5 fold more bone for histopathologic study. 2. The twin peak needle may produce less tissue hemorrhage and disruption.
Long-Term Follow-Up of Recombinant Human Erythropoietin (epoetin)Induced Pure Red Cell Aplasia (PRCA): A Comparison of Data Collected through Active and Passive Pharmacovigilance Efforts.
Authors: Bennett Charles L. , Carson Kenneth R. , McKoy June M. , Belknap Steven M. , Klinge Stephen A. , Evens Andrew M. , Raisch Dennis W. , Yarnold Paul R. , Kim Benjamin , Lyons Elizabeth A. , Kuzel Timothy K. , Tallman Martin S.
Published: October 2019

Background: Follow-up reports describing the long-term prognosis of 47 patients with epoetin induced PRCA have been confined to a single study from France, England, and Germany, investigated through active surveillance efforts. There is concern that reporting quality may differ between active versus voluntary reporting efforts.

Methods: Investigators from the Research on Adverse Drug reactions And Reports (RADAR) program compared clinical findings for epoetin-associated PRCA cases reported by voluntary surveillance efforts (n= 47) versus active surveillance (n= 47). (Verhelst et at, Lancet 2004). Recovery from PRCA was defined as transfusion independence, normalized reticulocyte count, and/or disappearance of anti-erythropoietin antibody.

Results: Baseline characteristics of age and gender were similar in both the MEDWatch and active surveillance groups. The two groups were similar with respect to median age similar (67 years) and gender (70%) and follow-up duration for patients who recovered (mean of 3 months). Median follow-up was shorter for passive surveillance patients who did not recover (6 versus 11 months). Recovery rates with immunosuppression differed (39.5% for MedWatch versus 21.6% for active surveillance).

Conclusions: Follow-up of active versus passive pharmacovigilance differed markedly for epoetin-associated PRCA cases in France, Germany, and the UK, in part due to differences in follow-up time. Active pharmacovigilance efforts should be supported by national health authorities.

Surveillance of Rx’ed patients Active N=37 (%) Passive N=38 (%) Patients with known follow-up (%) 100 100 Recovered w/transplant 16.2 18.4 Recovered w/Immunosuppression 62.2 42.1 No recovery 21.6 39.5
Active Site-Inactivated Recombinant Factor VIIa Blocks Shiga Toxin-Induced Upregulation of Tissue Factor Activity on Human Glomerular Endothelial Cells and Human Proximal Tubular Epithelial Cells.
Authors: Nestoridi Eirini , Kushak Rafail , Ingelfinger Julie R. , Grabowski Eric F.
Published: October 2019

Post-diarrheal hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in children, is most often caused by Shiga toxin (Stx)-producing Escherichia coli infection. The most widely recognized tissue damage in HUS occurs within the kidney, most prominently with local thrombosis and platelet-fibrin accumulation, suggesting activation of the coagulation cascade. We have shown that human proximal tubular epithelial cells (HK-2 cells) exposed to Stx-1 augment their constitutive surface tissue factor (TF) activity by 3.2-fold, and that Stx-1 treatment following TNF-alpha exposure enhances TF activity by 2.7-fold on human glomerular endothelial cells (HGECs). We investigated the possibility of modulating this cellular response to Stx-1 by blocking the TF pathway with active site-inactivated recombinant factor VIIa (irFVIIa), which retains the ability to bind TF, but is enzymatically inactive, thereby inhibiting thrombin formation. Moreover, its action is limited to exposed, functional TF and does not significantly prolong the bleeding time in vivo. Monolayers of both HK-2 cells and TNF-alpha-activated HGECs were incubated at 37°C with Stx-1. Functional TF was determined chromogenically as the production rate of factor Xa. The use of 60 nM irFVIIa completely abrogated the enhanced activity of TF caused by 4 hours incubation with 10 pM Stx-1 of TNF-alpha-activated HGECs (N = 4; Figure 1). On HK-2 cells, irFVIIa (60 – 200 nM) also effectively reversed the observed TF upregulation after exposure to 1 nM Stx-1 for 22 hours as well as blocked the constitutive TF expression which is seen with this cell type (N = 3; Figure 2). The TF pathway may prove to be of major importance in the pathogenesis of HUS, and direct targeting of this pathway with irFVIIa might provide a novel therapeutic approach to this disease.
Is the Risk of Breast Cancer Increased after Hematopoietic Cell Transplantation?.
Authors: Friedman Debra L. , Leisenring Wendy , Schwartz Jeffrey , Deeg H. Joachim
Published: October 2019

Survivors of hematopoietic stem cell transplantation (HSCT) are at risk for second malignant neoplasms (SMN). As secondary breast cancer is now reported in excess among survivors of a variety of cancers treated with conventional chemoradiotherapy, we sought to explore the risk among HSCT survivors. Among 4,805 allogeneic transplant recipients treated at the Fred Hutchinson Cancer Research Center between November, 1969 and April, 2003, who survived at least 100 days post transplant, there were 27 females and 1 male with secondary breast cancer, representing the largest single diagnostic group of secondary solid tumors post transplant, following cancers of the skin and mucosa. Confining the analysis to the 2,042 female HSCT recipients, the cumulative incidence of secondary breast cancer at 25 years was 5.4% (95% Confidence Interval [CI]: 3.0%, 8.9%). The risk was twice that of the general population, using US SEER data for expected values (Age and race adjusted standardized incidence ratio = 2.3; 95% CI = 1.5, 3.3). The time to second breast cancer from transplant was 1-24.8 (median 9.3) years. In Cox regression, using time from transplant as the time scale, risk increased with increasing age at time of transplant, with hazard ratios [HR] of 12.6 (95% CI 1.6,97.9) for patients 18–39 years and 54.7 (95% CI 6.6,450.8) for those ≥ 40 years. However, when age was used as the time scale, this age effect was no longer seen (HR: 0.56; 95% CI: 0.06,5,3). We also investigated the role of preparatory regimen and graft versus host disease (GVHD) and found no associations with increased breast cancer risk. Specifically, use of TBI in the conditioning regimen (HR: 1.9; 95% CI: 0.8,4.7), type of alkylating agent used in the conditioning regimen (Busulfan HR: 2.0; 95% CI: 0.1,32.0; Cytoxan HR: 1.3 95%; CI: 0.2,10.5; Busulfan plus Cytoxan HR: 0.8; 95% CI: 0.1,6.7), donor type (related versus unrelated; HR: 1.3; 95% CI 0.5,3.2), nor acute (HR: 1.2; 95% CI: 0.5,2.6) or chronic GVHD (HR: 1.5; 95% CI: 0.7,3.3) increased risk for secondary breast cancer. The only treatment-related risk factor identified was the use of single fraction (versus fractionated) TBI, where the hazard ratio was 4.8 (95% CI: 1.5,16.2). Among 1,015 100+ day survivors of autologous transplants, excluding those transplanted for primary breast cancer, there were 2 female and 1 male breast cancer reported. These data suggest that, in contrast to the risk reported following conventional chemoradiotherapy, transplant preparatory regimens do not significantly increase risk for secondary breast cancer. In addition, GVHD, which is a risk factor for a number of SMNs following HSCT, does not appear to increase the risk for secondary breast cancer. Risk factors other than therapy should therefore be investigated to better understand the increased risk of breast cancer following HSCT and to develop appropriate preventive strategies.