Autologous Heterotopic Transplant Of Peripheral Blood complementary of Neoadjuvant Chemotherapy in Solid Tumors

POSTER (1)Interdoctors (Montevideo, UY); (2)Interdoctors-France (Paris, FR); (3)Interdoctors-Argentina (Buenos Aires, AR); (4)Interdoctors-Uruguay (Montevideo, UY) 1096 interdoctors@live.com
Background Autologous bone marrow transplantation was proposed as a procedure of cancer immunotherapy based in the graft-versus host effect. In that model, graft/receptors were not conditioned to generate immunity responses, and frequently, the patients were immune-suppressed. Over the past few years, our team has developed an Autologous Heterotopic Transplant Of Peripheral Blood (AHTOPB), a procedure including a strong conditioning of graft/receptors to elicit antitumoral responses in the frame of a an Autologous Hemoderivative Cancer Vaccine (AHCV). Methods The state of art in the configuration of AHCV, including AHTOPB, is reported here for the first time: 1. Week 1 to 4: Patient conditioning with drugs that switch the malignancy-induced-tumor-progressive conditioning of the systemic microenvironment (neoangiogenesis/immune-tolerance). 2. Week 5 to 8: Transplant, ex vivo conditioned, to generate a subcutaneous/intramuscular site where injections of specific antitumoral immunization do not elicit tolerogenic responses. 3. Finally, thermostable autologous plasma fraction, previously reported as a fraction containing antigens released from tumors and conserved inside complexes of thermostable stress shock proteins, was injected in the prepared immunogenic site as prime immunization using chemotherapy as a booster. The evidence in independent reports for each step of this configuration is revisited. Patients: From clinical trials performed to test AHTOPB and previously published, control and treated patients, accomplishing the inclusion and exclusion criteria, were selected and the assessments statistically analyzed. Inclusion criteria: Primary solid tumors, Pancreas, Colorectal, NSCLC, Ovary, Prostate and Breast; non-resectable; M1; treated with standard neoadjuvant chemotherapy; tumor growth in progression according RECIST; PS 2 according ECOG scale. Exclusion criteria: co-morbidity in treatment; brain metastasis; surgery not performed; less than 3 months of neoadjuvant treatment. For each primary localization, patients were included in 2 groups: G1, treated with the standard chemotherapy and G2, treated with AHTOPB and Chemotherapy. Assessment: Overall Survival. Monthly, toxicities 3 (CTCAE) and number of cases in Stable Disease according RECIST. At time of diagnosis and post-treatment surgery, Lymphocyte Proliferation Assay (Thymidine-H3), Angiostatin and VEGF, T-Reg (ELISA) and Activated dendritic cells (IHC) were evaluated in tumor extracts; same laboratory tests in peripheral blood samples, post systemic treatment. Results Shown in Table Conclusions AHTOPB last configuration confirms safe antitumoral effect, improving the neoadjuvant activity of chemotherapy in different advanced solid tumors, stimulating future developments of this procedure addressed to its activity in the biological response. SY S T E M I C C O N D I T I O N I N G SITE CONDITIONING PRIME I M M U N I Z AT I O N BOOSTER
Pancreas 6.4 2.0 Colorectal 20.4 4.2 NSCLC 9.6 1.2 Ovary 50.0 6.2 Prostate 51.4 24 6.6 Breast 26.8 4.1

Lasalvia-Prisco E (1), Goldschmidt P (2), Galmarini F (3), Vzquez J (4), Lasalvia-Galante E (4), Cucchi S (4)

G1

14

18

14

14

G2

16

10.2 2.0

20

32.7 3.7

15

16.1 1.1

10

61.2 4.9

14

10

84.2 20 4.8

14

34.2 5.0

TX StD 3

OS mean SD

OS TX StD mean 3 SD

TX 3

StD

OS mean SD

OS TX n StD mean 3 SD

OS TX n StD mean n 3 SD

OS TX StD mean 3 SD

n: Number of cases; StD: Stable Disease (n);

OS: Overall Survival (months); TX: Toxicities CTCAE (n)

p<0.05

SAMPLES

VEGF

ANGIOSTATIN

T-Regulatory cells

Activated Dendritic Cells

Lymphocyte Proliferation Assay

Tumor extract at DB Mean SD Tumor extract at TS Mean SD PB Mean SD n Group

100 8.1 105 9.8 118 10.8 78 G1

100 7.7 60 7.2 74 6.7 79 G2

100 9.0 96 6.1 76 3.2 78 G1

100 8.4 210 2.6 178 8.4 79 G2

100 9.6 112 8.8 132 9.2 78 G1

100 9.5 48 1.6 52 2.5 79 G2

100 10.2 32 2.2 30 2.4 78 G1

100 9.7 148 7.8 126 6.9 79 G2

100 6.2 98 8.0 102 7.6 78 G1

100 5.4 305 13.1 201 11.0 79 G2

DB: Time of Diagnosis Biopsy; TS: Time of Tumor Surgery; PB: Post-Treatment Peripheral Blood Results expressed as % of mean value at DB p< 0.05

The hypothesis: The tolerogenic and polyvalent biological response is a leading phenomenon of malignancy; switching it is a strategic component of cancer treatments in order to avoid the progression or recurrence.
Peer Review reported Background
1. Autologous Induction of tumor fibrogenesis (Induccin autloga de fibrogenesis tumoral) (Autologous Induction of tumor fibrogenesis). Lasalvia E, Cucchi S, DeStefani E, Deneo H, Fierro L, Mechoso B, Larraaga J, Vzquez J. Neoplasia (Spain), 1995; (1):5-10 2. Anti-metastatic effect of a blood fraction from cancer patients. Lasalvia E, Cucchi S, Carlevaro T, Vzquez J, Riotorto R, Fierro L. Proc 31st ASCO Annual Meeting, May 1995: Abs 730 3. Serum Markers Variation Consistent with Autoschizis Induced by Ascorbic AcidMenadione in Patients with Prostate Cancer. Lasalvia-Prisco E, Cucchi S, Vzquez J, Lasalvia-Galante E, Golomar W, Gordon W. Medical Oncology, 2003; 20 (1): 45-52 4. Antitumoral Effect of a Vaccination Procedure with an Autologous Hemoderivative. Lasalvia-Prisco E, Cucchi S, Vzquez J, Lasalvia-Galante E, Golomar W, Gordon W. Cancer Biology & Therapy, 2003; 2 (2): 155-160 5. Breast Cancer: Autologous Immunogenicity Elicited by Chemotherapy. Lasalvia E, Cucchi S, Vzquez J, Lasalvia-Galante E, Golomar W, Gordon W. Proc. 39th ASCO Annual Meeting, May 2003: Abs 746 6. Insulin-induced Enhancement of Antitumoral Response to Methotrexate in Breast Cancer Patients. Lasalvia-Prisco E, Cucchi S, Vzquez J, Lasalvia-Galante E, Golomar W, Gordon W. Cancer Chemother Pharmacol, 2004; 53: 220-224 7. Colorectal Cancer: Autologous Immunogenicity in Chemotherapy (5FU) Pre-Treated Patients. Lasalvia-Prisco E, Cucchi S, Vazquez J, Lasalvia-Galante E, Golomar W, Cotto F, Otero R. Proc 40th ASCO Annual Meeting, May 2004: Abs 2593 8. Breast Cancer: Updated Vaccination with an Autologous Hemoderivative in changing Tumor Antigen Library. Garcia-Giralt E, Lasalvia-Prisco E, Cucchi S, Vazquez J. Proc 40th ASCO Annual Meeting, May 2004: Abs 2594 9. Breast Cancer: Role of Tumor Associated Antigens and Regulatory Cells [CD4+CD25+] as Targets of the Immune Response elicited by an Antiprogressive Autologous Hemoderivative Vaccine. Garcia-Giralt E, Lasalvia-Prisco E, Cucchi S, Vazquez J, Lasalvia-Galante E, Golomar W. Proc 41ST ASCO Annual Meeting, May 2005: Abs 2589 10. Colorectal Cancer: Comparative effects of an Autologous Hemoderivative Vaccine with a CEA vaccine and/or an autologous [CD4+CD25+] vaccine. Lasalvia-Prisco E, Garcia-Giralt E, Cucchi S, Vazquez J, Lasalvia-Galante E, Golomar W. Proc 41ST ASCO Annual Meeting, May 2005: Abs 2596 11. Advanced Colon Cancer: Antiprogressive Immunotherapy Using an Autologous Hemoderivative. Lasalvia-Prisco E, Garcia-Giralt E, Cucchi S, Vzquez J, Lasalvia-Galante E, Golomar W. Medical Oncology, 2006; 23 (1): 91-104 12. Advanced Breast Cancer: Antiprogressive Immunotherapy Using a Thermostable Autologous Hemoderivative. Lasalvia-Prisco E, Garcia-Giralt E, Cucchi S, Vzquez J, Lasalvia-Galante E, Golomar W. Breast Cancer Res Treat, 2006; 100: 149-160 13. Prostate cancer: autologous immunotherapy optimized by indoleamine- 2,3-dioxygenase (IDO)-inhibitor as immune-tolerance breaker. Lasalvia-Prisco E, Garcia-Giralt E, Cucchi S, Larraaga J. Proc 42nd ASCO Annual Meeting, May 2006: Abs 12509 14. Ovarian cancer: autologous immunotherapy optimized by remote adjuvancy of a silicate-induced granuloma. Garcia-Giralt E, Lasalvia- Prisco E, Cucchi S, Lasalvia-Galante E, Vazquez J, Golomar W, Vincent JP. Proc 42nd ASCO Annual Meeting, May 2006: Abs 12515 15. Prostate cancer: vaccine Sentinel Immunized Node (SIN) target for adjuvant locoregional chemotherapy in autologous vaccine. Lasalvia-Prisco E, Garcia-Giralt E, Cucchi S et al. Proc 43rd ASCO Annual Meeting, June 2007: Abs 13500 16. Breast cancer: draining lymph node of vaccination site targeted by adjuvant GM-CSF in an autologous vaccine. Garcia-Giralt E, Lasalvia-Prisco E, Cucchi S et al. Proc 43rd ASCO Annual Meeting, June 2007: Abs 13506 17. Immunotherapeutic Site in Breast Cancer: a new methodology for the Autologous Thermostable Hemoderivative Cancer Vaccine (ATH-CV). Garcia-Giralt E, Lasalvia-Prisco E, Cucchi S et al. European Institute of Oncology, 9th Milan Breast Cancer Conference, June 20-22, 2007, Innovation in Cancer Research, Chair: Veronesi, U - Goldhrish, A, Coord: Cinieri S, Colleoni M, Zurrida S. Section I, I-1 18. Conditioning of vaccine sentinel lymph node as adjuvant of Autologous Hemoderivative Breast Cancer Vaccine. Lasalvia-Prisco E, GarciaGiralt E, Cucchi S, et al. Poster Presentation. 14th European Cancer Conference. Barcelona, Spain, September 2007: Poster 804 19. Advanced Ovarian Cancer: Vaccination Site Draining Lymph Node as Target of Immunomodulative Adjuvant in Autologous Cancer Vaccine. Lasalvia-Prisco E, Garcia-Giralt E, Cucchi S et al. Biologics: Targets & Therapy, October 2007; 1 (2):173-181 20. Erythrocytes as autologous-vaccine carriers targeted in breast cancer sentinel lymph node. Garcia-Giralt E, Lasalvia-Prisco E, Cucchi S et al. ASCO 2008. J Clin Oncol 26: 2008 (May 20 suppl; Abs 14018) 21. Erythrocytes as Autologous-Vaccine Carriers in Advanced Prostate Cancer. Lasalvia-Prisco E, Garcia-Giralt E, Lasalvia-Galante E et al. ASCO 2008 J Clin Oncol 26: 2008 (May 20 suppl; Abs 14017) 22. Randomized phase II clinical trial of Chemo-immunotherapy in Advanced Non-Small Cell Lung Cancer. Lasalvia-Prisco E, Garcia-Giralt E, Vzquez J et al. Biologics: Targets & Therapy, 2008; 2 (3): 555561 23. From Classic Autohemotherapy to Autologous Hemoderivative Cancer Vaccine through a drug and drug-carrier Immunomodulatory Adjuvant System. Lasalvia-Prisco E, Garcia-Giralt E, Vzquez J, Cucchi S, Lasalvia-Galante E, Larraaga J. Poster Presentation. ERLICH II, 2nd World Conference, Nurnberg, Germany, Oct 2008 24. Autologous Heterotopic Transplant of Peripheral Blood as a Competitive Site Against the Tolerogenic Activity of Human Malignant Tumors. Cochair and Presenter at Session 1-5: Tumor Immunology, 1st Annual World Congress of Immunodiseases and Therapy, Beijing, China, May 2010 25. Autologous Heterotopic Transplant of Peripheral Blood (AHTOPB) as conditioner of the Antitumoral Biological Response. Lasalvia-Prisco E, Garcia-Giralt E, Cucchi S, Vazquez J. Poster Presentation. XXXIII World Congress of the International Society of Hematology, Jerusalem, Israel, October 2010 26. Pancreatic cancer: Immunotherapy and Antiangiogenesis elicited with conditioned Autologous Heterotopic Transplant Of Peripheral Blood. Lasalvia-Prisco E, Uruguay; Garcia-Giralt E, France; Goldschmidt P, France; Cucchi S, Uruguay; Vzquez J, Uruguay; Galmarini R, Argentina. Poster Presentation. ESMO 13th World Congress on Gastrointestinal Cancer, Barcelona, Spain, June 2011 27. Mechanism of action for antitumoral activity of autologous heterotopic transplant of Peripheral Blood in Non Small Cell Lung Cancer. LasalviaPrisco E, Garcia-Giralt E, Goldschmidt P, Cucchi S, Vazquez J, Galmarini F. Poster Presentation. 2011 European Multidisciplinary Cancer Congress-ECCO 16-ESMO 36-ESTRO 30. Stockholm, Sweden, September 23-27, 2011
COMMENTS ABOUT THIS SUBJECT Commentary: A New Twist on Autologous Cancer Vaccines. Leisha A. Emens MD. PhD, Assistant Professor of Oncology and Director of the Tumor Immunology and Immunotherapy Laboratory in Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins University, Baltimore, Maryland. Cancer Biology & Therapy, March/April 2003; 2 (2):161-163

PROCEDURE

Immunization-site sentinel lymph node

Autologous Heterotopic Transplant Of Peripheral Blood

RATIONAL
Systemic Switch of malignancy-induced tolerogenic conditioning T-Reg Cox2-I Neutrophil Activation Plasmin Angiostatin Deletion IDO T-Reg Recruitment Dendritic cell Angiostatin Infiltration

TECHNOLOGY
SYSTEMIC CONDITIONING

PRIME IMMUNIZATION

BOOSTER IMMUNIZATION

Cox2- Inhibitor
G-CS F

Cyclophosphamide
SH donor

SITE CONDITIONING Autologous Heterotopic Transplant Of Peripheral Blood

Blood extraction Antigenic Fraction Carrier

Chemotherapy Re-injection

Local site conditioning to generate no-tolerogenic immunity response Platelet / Neutrophil Thrombin PAR Activation Sequential degranulation VEGF and ANGIOSTATIN RED CELL / HEM PMBC release cytokines Gamma If- IL2-IL12

Apoptosis of stressed tumor cells Free in Thermostable plasma fraction

Tumor antigens released to the blood Complexed in Stress (Heat) Shock Proteins

Extraction

Activation

Re-injection

GRANULOMA EFFECT

LYMPH NODE OF IMMUNIZATION SITE Increase Activated Dendritic Cells Decrease T-Reg Cells

Se le c t e d

R e f e r e n ces

1 Metronomic cyclophosphamide regimen selectively depletes CD4+ CD25+ regulatory T cells and restores T and NK effector functions in end stage cancer patient. Ghiringhelli F, et al. Cancer Immunol Immunother, 2006; 56: 641 2 CD4+CD25+ regulatory T cells suppress tumor immunity but are sensitive to cyclophosphamide which allows immunotherapy of established tumors to be curative. Ghiringhelli F, et al. Eur J Immunol, 2004; 34: 336 3 The Immune Tolerance of Cancer is Mediated by IDO That is Inhibited by COX-2 Inhibitors Through Regulatory T Cells. Lee SY, et al. J Immunother, 2009; 32: 22 4 Cyclooxygenase-2 Inhibitor Enhances the Efficacy of a Breast Cancer Vaccine: Role of IDO. Basu GD. J Immunol, 2006, 177: 2391 5 Specific Inhibition of Cyclooxygenase 2 Restores Antitumor Reactivity by Altering the Balance of IL-10 and IL-12 Synthesis. Stolina M, et al. J Immunol, 2000, 164: 361 6 Neutrophil Activation and Hemostatic Changes in Healthy Donors Receiving Granulocyte Colony-Stimulating Factor. Falanga A, et al. Blood, 1999; 93(8): 2506 7 The mechanism of cancer-mediated conversion of plasminogen to the angiogenesis inhibitor angiostatin. Gately S, et al. Proc. Natl. Acad. Sci. USA, 1997; 94: 10868 8 In vivo Generation of Angiostatin Isoforms by Administration of a Plasminogen Activator and a Free Sulfhydryl Donor: A Phase I Study of an Angiostatic Cocktail of Tissue Plasminogen Activator and Mesna. Soff GA. Clin Cancer Res, 2005; 11(17): 6218 9 In Vitro and In Vivo Induction of Antiangiogenic Activity by Plasminogen Activators and Captopril. Merchan JR. J Natl Cancer Inst, 2003; 95(5): 388

10 Anti-angiogenesis therapy can overcome endothelial cell anergy and promote leukocyte-endothelium interactions and infiltration in tumors. Dirkx AEM, et al. FASEB J, 2006; 20: 621 11 Thrombin and Platelet Activation. Brass LF. Chest 2003; 124(3): 18S 12 Proteinase-activated receptor-2: expression by human neutrophils. Howells GL, et al. Journal of Cell Science, 1997; 110: 881 13 Generation and role of angiostatin in human platelets. Jurasz P, et al. Blood, 2003; 102: 3217 14 Angiogenesis is regulated by a novel mechanism: pro- and antiangiogenic proteins are organized into separate platelet granules and differentially released. Italiano Jr JE, et al. Blood, 2008; 111 (3): 1227 15 Autologous Red Blood Cells Potentiate Antibody Synthesis by Unfractionated Human Mononuclear Cell Cultures. Rugeles MT, et al. Scand J Immunol, 26(2): 119 16 Enhancing Effects of Autologous Erythrocytes on Human or Mouse Cytokine Secretion and IL-2R Expression. Kalechman Y, et al. Cell Immunol, 1993; 148: 114 17 Inflammation and Host Resistance against tumours I V. Fauve RM, Hevin MB, Fontan E, et al. Ann Immunol (Paris), 1977; 128C: 923 & 1982; 133C (1):133 & Int J Cancer, 1988; 15; 42(2): 267 & 1990; 46(3): 533 18 Sentinel node mapping identifies vaccine-draining lymph nodes with tumor-specific immunological activity. Chin CS and Bear HD. Ann Surg Oncol, 2002; 9 (1): 94 19 Pathologic Complete Response to Neoadjuvant Chemotherapy of Breast Carcinoma Is Associated with the Disappearance of Tumor-Infiltrating Foxp3+ Regulatory T Cells. Ladoire S, et al. Clin Cancer Res, 2008; 14(8): 2413

20 Carcinoembryonic Antigen as a Target for Therapeutic Anticancer Vaccines: A Review. Berinstein NL. J Clin Oncol, 2002; 20: 2197 21 Immunology and Immunotherapy Approaches for Prostate Cancer. Elkord E. Prostatic Dis, 2007; 10(3): 224 22 Soluble tumor-associated antigens in cancer detection, prevention and therapy. Zusman I. Med Sci Monit, 2004; 10(12): RA317 23 Thermostable antigens of malignant tumors and normal tissues of experimental animals. Lomakin MS, et al. Biull Eksp Biol Med, 1978; 85(6): 726 24 Two-dimensional electrophoretic proteome study of serum thermostable fraction from patients with various tumor conditions. Goufman EI, et al. Biokhimiya, 2006; 71(4): 445 25 Immunotherapy of Tumors with Autologous Tumor-Derived Heat Shock Protein Preparations. Tamura Y, et al. Science, 1997; 278: 117 26 Thermo-stable tumor-associated antigens in the serum of tumor-bearing animals and in tissue cultures of malignant tumors. Lomakin MS, et al. Biull Eksp Biol Med, 1980; 89(4): 452 27 Heat Shock ProteinPeptide Complexes, Reconstituted In Vitro, Elicit Peptide-specific Cytotoxic T Lymphocyte Response and Tumor Immunity. Blachere N, et al. J Exp Med, 1997; 186(8): 1315 28 Subcutaneous Administration of Carrier Erythrocytes: Slow Release of Entrapped Agents. DeLoach JR and Corrier DE. Biotechnology and Applied Biochemistry, 1998; 10(4): 359

29 Applications of carrier erythrocytes in delivery of biopharmaceuticals. Hamidi M, et al. Journal of Controlled Release, 2007; 118: 145 30 Tumor immunogenicity is determined by the mechanism of cell death via induction of heat shock protein expression. Melcher A, et al. Nature Medicine, 1998; 4(5): 581 31 Chemotherapy: induction of stress responses. Tiligada E. EndocrineRelated Cancer, 2006; 13: S115 32 Chemotherapy Delivered After Viral Immunogene Therapy Augments Antitumor Efficacy Via Multiple Immune-mediated Mechanisms. Fridlender ZG, et al. Molecular Therapy, 2010; 18(11): 1947 33 The mouse immune response to carrier erythrocyte entrapped antigens. Murray AM. Vaccine, 2006; 24: 6129 34 Induction of Tumor Cell Apoptosis In Vivo Increases Tumor Antigen CrossPresentation, Cross-Priming Rather than Cross-Tolerizing Host Tumor-Specific CD8 T Cells. Nowak AK, et al. J Immunol, 2003; 170: 4905

Network of Research & Development of Medical Procedures Telemedical Organization, American Medical Intelligence. Florida, USA

interdoctors
www.cancerinterdoctors.com interdoctors@live.com

European Group for Blood and Marrow Transplantation - EBMT 2012 - Geneva, Switzerland, 1 - 4 April 2012