Condition category
Cancer
Date applied
30/10/2007
Date assigned
31/10/2007
Last edited
31/10/2007
Prospective/Retrospective
Retrospectively registered
Overall trial status
Completed
Recruitment status
No longer recruiting

Plain English Summary

Not provided at time of registration

Trial website

Contact information

Type

Scientific

Primary contact

Prof Michael Schmitt

ORCID ID

Contact details

Universitatsklinikum Ulm
Robert-Koch-Str.8
89081 Ulm
Ulm
89081
Germany
michael.schmitt@uniklinik-ulm.de

Additional identifiers

EudraCT number

ClinicalTrials.gov number

Protocol/serial number

N/A

Study information

Scientific title

Acronym

RHAMM peptide vaccination

Study hypothesis

Primary study aim:
Proof of the safety and feasibility of a vaccination with this particular peptide in patients with haematological malignancies.

Primary endpoint:
1. Frequency of Severe Adverse Events (SAE)
2. Severity of SAE
3. Timepoints and correlations to the study medication of the SAE

Secondary aims of the study:
1. Induction of a specific T cell immune reponse to RHAMM/CD168
2. Assessment of the influence of the peptide vaccination on the remission status of the present haematological malignancy

Secondary endpoints:
1. Frequency of RHAMM specific T Lymphocytes
2. Remission criteria

Ethics approval

Ethics approval received from the Ethics committee of the University of Ulm (Germany) on the 24th September 2004 (ref: UL-RHAMM-1).

Study design

Interventional, non-randomised, non-controlled, pilot study

Primary study design

Interventional

Secondary study design

Non randomised controlled trial

Trial setting

Hospitals

Trial type

Treatment

Patient information sheet

Condition

Acute Myeloid Leukaemia (AML), Myelodysplastic Syndrome (MDS) or Multiple Myeloma (MM)

Intervention

300 µg RHAMM R3 peptide emulsified with the incomplete Freund's adjuvant on day 3 as well as Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) on days 1 - 5 was administered four times subcutaneously at a biweekly interval.

Timepoints:
Pre examination: week 0
RHAMM-R3 vaccinations: weeks 1,3,5,7 with intermediate examinations
Final examination: week 10

Intervention type

Drug

Phase

Phase I/II

Drug names

Receptor for Hyaluronan-Mediated Motility (RHAMM [CD168]) peptide, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF)

Primary outcome measures

For all patients the BM blood was analysed before and after vaccination using microscopy and standard Fluorescence Activated Cell Sorter (FACS) analysis. Patients with MM were also examined for quantitative immunoglobulins and quantitative free light chains in serum and urine. The frequency of erythrocyte and platelet transfusions and the course of differential blood count were documented.

Response criteria were as following:
For patients with AML, the criteria by the World Health Organization (WHO) and the International Working Group (IWG) were followed as specified:
Complete Response (CR): reduction of blasts in the BM blood to less than 5%, in peripheral blood count: haemoglobin greater than 11 g/dl, neutrophils 1,500/mm^3 or more, platelets 100,000/mm^3 or more
Partial Response (PR): reduction of blasts in the BM blood of more than 50%, in peripheral blood count: haemoglobin greater than 11 g/dl, neutrophils 1500/mm^3 or more, platelets 100,000/mm^3 or more
Stable Disease (SD): no CR or PR
Progressive Disease (PD): increase of blasts in the BM blood by more than 50% or increase of WHO-classification or progress of transfusion requirements

For patients with MDS, the criteria by the WHO and the IWG were followed as specified:
CR: a complete response was defined as a normocellular with less than 5% blasts with normal maturation of all cell lines, with no evidence of dysplasia, in peripheral blood count: haemoglobin greater than 11 g/dl, neutrophils 1,500/mm^3 or more, platelets 100,000/mm^3 or more
PR: blasts decreased by 50% or more over treatment or a less MDS WHO classification than pretreatment, Haematological Improvement (HI): an improvement was defined as a decrease of at least of 50% in transfusion requirements, together with at least an improvement of one ot two cell lineages of the peripheral cell counts but not enough to qualify for a PR
SD: failure to achieve at least a HI, but no evidence of progression for at least two months
PD: increase of blasts in bone-marrow of more than 50% or increase of WHO-classification or progress of transfusion requirements

For patients with MM, the International Uniform Response Criteria according to Durie et. al. were applied:
Stringent Complete Remission (sCR): CR as defined below plus normal free light chain ratio and absence of clonal cells in the BM by immunohistochemistry or immunofluorescence
CR: negative immunofixation in the serum and urine
Very Good Partial Response (VGPR): serum and urine M-protein detectable by immunofixation but not on electrophoresis or 90% or greater reduction in serum M-protein plus urine M-protein less than 100 mg per 24 hours
PR: greater than or equal to 50% reduction of serum M-protein and reduction in 24-hour urinary M-protein by greater than or equal to 90% or to less than 200 mg per 24 hours
SD: no CR or PR
PD: increase of free light chains in serum or urine or of clonal plasma cells in bone-marrow of more than 25%

Secondary outcome measures

1. Assessment of toxicity of R3-peptide vaccination:
Side effects were documented according to Common Terminology Criteria for Adverse Events v3.0 (CTCAE). Before and three weeks after the fourth vaccination physical examination, body weight, ECOG performance score, laboratory tests (kidney and liver function tests, electrophoresis, electrolytes, C-Reactive Protein [CRP], Lactate Dehydrogenase [LDH], and coagulation tests), chest x-ray, echocardiography, electrocardiography, urine analysis, abdominal sonography and bone-marrow aspiration was performed. For patients with MM additionally quantitative immunoglobulins and quantitative assessment of free light chains in serum and urine were tested. Before each vaccination physical examination, laboratory tests (White Blood Cell [WBC] count, differential blood count, kidney and liver function tests, electrolytes, CRP, LDH, coagulation tests and urine analysis) were performed. To detect autoimmune reactions, we measured Thyroid Stimulating Hormone (TSH), free Triiodothyronine (fT3), free Thyroxine (fT4), as well as Antinuclear Antibody (ANA), CRP and Blood Sedimentation rate (BSG).

2. Interferon (IFN)-gamma and Granzyme B Enzyme-Linked Immunosorbent Spot (ELISpot) assays:
IFN-gamma and granzyme B ELISpot assays were performed as previously described to determine specific lysis of RHAMM (peptide) positive target cells according to the manufacturer's instructions (BD, San Diego, USA). We participated in an inter-laboratory test for ELISpot assays.

3. Tetramer staining:
The frequency of R3 specific CD8+ T lymphocytes was determined after eight days Mixed Lymphocyte Peptide Culture (MLPC) by staining with anti-CD8 antibody and HLA-A2/R3 tetramer R-Phycoerythrin (PE). HLA-A2/R3 tetramer PE was synthesised at the Lausanne Branch of the Ludwig Institute for Cancer Research. CD8+ T lymphocytes (0.5 - 1 x 10^6) stimulated with irradiated CD8- Antigen-Presenting Cells (APCs) in the presence of the R3 peptide were stained with HLA-A2/R3 tetramer PE 1 µg per test with respect to the peptide-Major Histocompatibility Complex (MHC) class I component in the dark and incubated for 40 minutes at room temperature. Thereafter, for four-color staining, 10 µl CD8 Peridinin Chlorophyll Protein (PerCP), 10 µl CD45RA Fluorescein Isothiocyanate (FITC) and 4 µl CCR7 APC (BD, Heidelberg, Germany) were added at 4°C for 20 minutes in the dark. As for six-color staining, the cells were stained with 1 µg HLA-A2/R3 tetramer PE and HLA-A2/WT1 tetramer PerCP per test with respect to the peptide-MHC class I component in the dark and incubated for 40 minutes at room temperature. Thereafter 5 µl CD8 APC-Cy7, 5 µl CD45RA APC (Invitrogen, Caltag, CA, USA), 10 µl CCR7 PE-Cy7, 10 µl CD27 FITC or CD28 FITC (BD, Heidelberg, Germany) were added at 4°C for 20 minutes in the dark. After washing once with Phosphate-Buffered Saline (PBS), stained cells were fixed with 1% formaldehyde (Sigma, Germany) and then analysed by flow cytometry. Whenever possible, at least 100,000 events were collected for analysis. Each sample was run with an appropriate isotype control to define the gate of positive cells. Analysis was performed on tightly gated lymphocytes to exclude dead cells and debris and on CD8+ T lymphocytes to evaluate responses to R3 peptide. Samples were defined as "tetramer positive" in case of an increase of specific R3-tetramer+/CD8+ T cells of more than 50% (if initial count was = 0.1%), or 25% increase (if initial count was greater than 0.1%). We participated in an inter-laboratory test for tetramer flow cytometry assays.

Overall trial start date

01/11/2004

Overall trial end date

01/02/2008

Reason abandoned

Eligibility

Participant inclusion criteria

1. Diagnosis of Acute Myeloid Leukaemia (AML), Myelodysplastic Syndrome (MDS) or Multiple Myeloma (MM)
2. AML: up to 25% blasts in the Bone Marrow (BM); MDS: up to 20% blasts in the BM; MM: partial remission (immunofixation still positive or immunoglobulins still detectable in the urine)
3. Human Leukocyte Antigen A2 (HLA-A2) expression
4. RHAMM-messenger Ribonucleic Acid (mRNA) expression
5. Karnofsky Index greater than or equal to 70 or Eastern Cooperative Oncology Group (ECOG) performance status 0 to 2
6. Aged greater than 18 years
7. At least one cycle of treatment with standard chemotherapy for this haematological malignancy preceding the peptide vaccination
8. Survival time at least 6 months
9. Sufficient renal function (creatinine and Blood Urea Nitrogen [BUN] less than threefold of the upper limit)
10. Sufficient liver function tests (Serum Glutamic Oxaloacetic Transaminase [SGOT]/Serum Glutamic Pyruvic Transaminase [SGPT] threefold of the upper limit)
11. Compliance of the patient
12. Informed consent must be obtained in written form

Participant type

Patient

Age group

Adult

Gender

Both

Target number of participants

21

Participant exclusion criteria

1. Allogeneic hematopoietic stem cell transplantation in the history planned
2. Central Nervous System (CNS) involvment, severe psychiatric disease
3. Severe partial or global respiratory failure (New York Heart Association [NYHA] stage greater than or equal to III)
4. Immunosuppressive therapy in the last 4 weeks
5. Pregnancy or breastfeeding
6. Females with no sufficient contraception
7. Contradictions against study therapeuticals (including galenic substances)
8. Severe infections
9. Simultaneous participation in another clinical study trial

Recruitment start date

01/11/2004

Recruitment end date

01/02/2008

Locations

Countries of recruitment

Germany

Trial participating centre

Universitatsklinikum Ulm
Ulm
89081
Germany

Sponsor information

Organisation

University Hospital Ulm (Universitatsklinikum Ulm) (Germany)

Sponsor details

c/o Professor R Marre
Robert-Koch-Str. 8
Ulm
89081
Germany

Sponsor type

Hospital/treatment centre

Website

http://www.uniklinik-ulm.de/

Funders

Funder type

Government

Funder name

This work was supported by generous grants from:

Alternative name(s)

Funding Body Type

Funding Body Subtype

Location

Funder name

The German José Carreras Leukemia Foundation (Germany)

Alternative name(s)

Funding Body Type

Funding Body Subtype

Location

Funder name

German Research Council (Deutsche Forschungsgemeinschaft [DFG]) (Germany)

Alternative name(s)

Funding Body Type

Funding Body Subtype

Location

Results and Publications

Publication and dissemination plan

Not provided at time of registration

Intention to publish date

Participant level data

Not provided at time of registration

Results - basic reporting

Publication summary

Publication citations

Additional files

Editorial Notes