A new immunotherapy strategy targeted CD30 in peripheral T-cell lymphomas: CAR-modified T-cell therapy based on CD30 mAb

Chimeric antigen receptor T-cell immunotherapy (CAR-T) has shown remarkable efficacy in treating tumors of lymphopoietic origin. Herein, we demonstrate an effective CAR-T cell treatment for recurrent and malignant CD30-positive peripheral T-cell lymphomas (PTCL) has been demonstrated. The extracellular fragment gene sequences of CD30 were obtained from tumor tissues of PTCL patients and cloned into a plasmid vector to express the CD30 antigen. The CD30 targeting single-chain antibody fragment (scFv) was obtained from CD30-positive monoclonal hybridoma cells, which were obtained from CD30 antigen immunized mice. After a second-generation of CAR lentiviral construction, CD30 CAR T cells were produced and used to determine the cytotoxicity of this construct toward Karpas 299 cells. The results of CD30 CAR T-mediated cell lysis show that 9C11-2 CAR T cells could significantly promote the lysis of CD30-positive Karpas 299 cells in both LDH and real-time cell electronic sensing (RTCA) assays. In vivo data show that 9C11-2 CAR T cells effectively suppress the tumor growth in a Karpas 299 cell xenograft NCG mouse model. The CD30 CAR T cells exhibited an efficient cytotoxic effect after being co-cultured with the target cells and they also exhibited a significant tumor-inhibiting ability after being intravenously injected into PTCL xenograft tumors; these observations suggest that the new CD30 CAR-T cell may be a promising therapeutic candidate for cancer therapy.

Brentuximab vedotin is the first target drug in the treatment of hodgkin lymphoma

Progress in the treatment of Hodgkin lymphoma are among the most significant achievements of oncology of our age. Nevertheless, an early relapse or refractory course of the disease account for approximately 15–20% of cases. It was this category of patients, in which the target drug brentuximab vedotin was successfully used for the first time. This is an anti-CD30 (a cell surface antigen) humanized monoclonal antibody conjugated via a protease-cleavable linker to the cytostatic agent monomethyl auristatin E, the potent tubulin inhibitor. This article describes the characteristics of CD30 antigen and a new preparation, as well as clinical data confirming its efficacy, the results of major reviews and research on the use of brentuximab vedotin in mono and combination therapy regimens at the different stages of treatment for Hodgkin lymphoma.

Use of CD30 expression in human malignant mesothelioma as a novel therapeutic target.

e18526 Background: CD30 antigen is important in the diagnosis of Hodgkin’s disease (HD) and anaplastic large cell lymphoma (ALCL). CD30 is a cytokine receptor that belongs to the tumor necrosis factor superfamily (TNFRSF8) and acts as a positive regulator of apoptosis. The expression of CD30 in malignant mesothelioma is unknown. Given the remarkable success of brentuximab vedotin, an antibody-toxin conjugate directed against CD30 antigen, in treating patients with relapsed HD and ALCL, we undertook a study to validate the incidence of CD30 expression in mesothelioma and to examine its clinical correlates. Furthermore, we performed studies using cultured mesothelioma and non-small cell lung cancer (NSCLC) cell lines to determine the expression and potential targeting of CD30 antigen in vitro. Methods: Patient mesothelioma tissue specimens (n=52) were examined for CD30 expression by immuno-histochemistry (IHC) with an anti-BerH2 antibody (Cell Marque) used for the diagnosis of HD. Staining was assessed by a thoracic pathologist. Mesothelioma (H28, H2052, H2452, 211H) and NSCLC (A549, H1299) cell lines were also grown in culture and examined for CD30 expression by FACS analysis and confocal microscopy using anti-CD30 antibody (Novus Biologicals). Cells were permeabilized with Triton X-100. Results: Positive CD30 expression was noted in six out of 52 total mesothelioma specimens. Five of the six positive tumors demonstrated epithelial histology and were scored as low or intermediate grade. Membrane-associated as well as diffuse cytoplasmic staining was observed. The percentage of tumor cells stained positive varied greatly. Interestingly, no high grade epithelial tumors were scored as CD30 positive. The remaining CD30 positive tumor was a high grade, biphasic metastatic tumor. Confocal analysis of both mesothelioma cultured cells localized CD30 antigen to the cell membrane. Flow cytometry of intact and permeabilized mesothelioma cells supported the idea that CD30 antigen was located on the cell-surface. Conclusions: Our studies validate the presence of CD30 antigen on the cell surface of epithelial-type mesothelioma and indicate that selected mesothelioma patients may derive benefit from brentuximab vedotin (Adcetris).

CD30 - the head of TNF-family… or a successful story of brentuximab vedotin

Hodgkin lymphoma and anaplastic large cell lymphoma are malignancies that highly express CD30 antigen on the cell surface. Both are generally curable by standard chemotherapy but refractory diseases and relapses are treatment problems. Brentuximab-vedotin is a labeled monoclonal antibody against CD30 and it is approved for the treatment of Hodgkin lymphoma relapsed after autologous stem cell transplantation and for relapsed anaplastic large cell lymphoma. This is the first drug approved for the treatment of Hodgkin lymphoma after 30 years.

Preclinical and Clinical Binding Properties, Internalization Kinetics, and Clinicopathological Activity of Brentuximab Vedotin (SGN-35): A Novel Antibody Drug Conjugate for Anaplastic Large Cell Lymphoma and Classical Hodgkin Lymphoma

Abstract 1789 Background: Brentuximab vedotin (SGN-35) is a novel anti-CD30 antibody conjugated to the cytotoxic drug monomethyl auristatin E (MMAE) designed to selectively target and kill CD30 expressing neoplasms. This agent has demonstrated antitumor activity in classical Hodgkin lymphoma and CD30-positive T cell lymphoma, yet the binding properties, internalization kinetics, and clinicopathological findings have not been described in tumor specimens derived from treated patients. Therefore, we investigated the activity of SGN-35 on a patient with cutaneous manifestations of systemic ALK-negative anaplastic large cell lymphoma (ALCL), and correlated these results with studies of the activity of SGN-35 on cultured CD30-positive cells lines. Methods and Results: First, we confirmed (using flow cytometry) that SGN-35 and the anti-CD30 antibody clone BerH83 do not compete for binding to CD30. Next, in Karpas 299 (ALCL cell line) and KM-H2 (cell line derived from Hodgkin lymphoma) cells, we quantitatively monitored CD30 expression and cell-surface SGN-35 over time (measured as antigen binding capacity [ABC]/cell), demonstrating maximum antigen expression at 24–48 hours of incubation with 15 μg/ml SGN-35 followed by decrease at 120 hours, consistent with SGN-35-induced internalization (Table). Similar studies were performed on sequential tumor biopsies from a 68 year old male with a 5-year history of ALK-negative ALCL relapsing after 4 prior systemic regimens and radiation, now presenting with multiple pink ulcerated tumorous lesions on his lower extremities as well as bone and nodal involvement. H&E and anti-CD30 immunohistochemical stains of both pretreatment and 24 hrs post-treatment (after the first dose of SGN-35 (1.8mg/kg)-ClinicalTrials.gov identifier NCT01026415) skin punch biopsies showed a dense dermal infiltrate of large CD30-positive neoplastic cells. Biopsy at 48 hrs post-treatment demonstrated numerous apoptotic cells. Antigen density experiments on the patient biopsy specimens and clinical findings correlated with these morphologic results. CD30 antigen density was highest on the patient's cells pretreatment biopsy (1.01 × 105 ABC units/cell) and then decreased after 24 hrs (7.83 × 104 ABC units/cell) and 48 hrs (5.08 × 104 ABC units/cell). Bound SGN-35 was greatest at 24 hrs (2.26 × 103 ABC units/cell) and decreased at 48 hrs (1.40 × 103 ABC units/cell) (Table). The corresponding measured blood concentrations of SGN-35 at 24 hr, 48 hr, and 21 days were 10 μg/mL, 7 μg/mL, and 1 μg/mL, respectively. No CD30 positive cells were present in the day 21 biopsy, precluding evaluation of CD30 antigen density and bound SGN-35. Clinically, the size and prominence of the skin lesions were reduced at day 21 post treatment and biopsy at this time point showed no morphologic or immunohistochemical evidence of the neoplastic population (pathologic complete remission). After the second dose of SGN-35 the patient achieved an 81% reduction in all radiographically measurable lesions (partial response) and achieved a radiographic and cutaneous complete remission after the 5th infusion of this agent. Conclusion: These data are the first to suggest in both cell lines and patient-derived tissues that the internalization kinetics of SGN-35 is rapid with resultant reduction in CD30 expression within the first 48 hours and concurrent apoptosis induction within the targeted cells. While the measured SGN-35 occupancy of CD30 binding sites in the patient is lower than with the cell lines, these data highlight the potential antitumor activity of this agent. The results from the single patient need to be confirmed in a cohort of patients. Nevertheless, such results imply that even with subsaturating occupancy of CD30, thousands of molecules of SGN-35 (each with approximately 4 molecules of MMAE) are likely to be internalized by and potentially kill each targeted cell and may be sufficient to yield pathologic remissions and clinical activity. Disclosures: Fromm: Seattle Genetics: Research Funding. McEarchern:Seattle Genetics: Employment. Kennedy:Seattle Genetics: Employment. Shustov:Seattle Genetics: Research Funding; Seattle Genetics: Membership on an entity's Board of Directors or advisory committees. Gopal:Seattle Genetics: Research Funding.

Therapies for Relapsed Hodgkin Lymphoma: Transplant and Non-Transplant Approaches Including Immunotherapy

Autologous stem cell transplant remains the standard of care for relapsed Hodgkin lymphoma (HL). Approximately 50% of patients with chemo-sensitive relapse will be cured with this approach. The optimal pretransplant salvage regimen is controversial, but less toxic combinations seem to be equivalent to more aggressive approaches. For patients with chemo-refractory disease at relapse and those failing autologous transplant, the long-term prognosis remains poor. New approaches such as reduced-intensity allogeneic transplant, monoclonal antibodies targeting the CD30 antigen, Epstein-Barr virus (EBV)-specific cytotoxic T-lymphocytes, and bortezomib are under investigation, but preliminary results are disappointing. New therapies are needed for patients with relapsed HL.