Background Systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS) are autoimmune disorders marked by the presence of pathogenic autoantibodies predominantly produced by B cells. Both conditions are chronic, incurable, and require lifelong treatment. Chimeric antigen receptor T-cell (CAR-T) therapy has emerged as a promising treatment for refractory hematological malignancies, with anti-CD19 CAR-T therapy demonstrating high efficacy in relapsed or refractory diffuse large B-cell lymphoma (DLBCL). The profound depletion of B cells induced by anti-CD19 CAR-T therapy suggests that this treatment may influence the clinical course of patients with DLBCL and concomitant SLE or APS. Materials and methods We report two clinical cases of patients diagnosed with both DLBCL and concomitant SLE and APS who underwent anti-CD19 CAR-T therapy at our institution. We describe the lymphoproliferative disease course as well as the serological evolution and longitudinal cytokine profiles measured before and after CAR-T therapy. Results In both cases, CAR-T therapy resulted in complete and long-lasting metabolic remission of the lymphoproliferative disorder. Concurrently, autoantibody titers stabilized, permitting stepwise de-escalation of the immunomodulatory treatment. Conclusion These cases demonstrate that CAR-T therapy for DLBCL is feasible and appears safe in patients with concomitant autoimmune disease but requires close monitoring and interdisciplinary management. These findings support further preclinical and clinical investigation of CAR-T therapy as a potential therapeutic approach in patients with DLBCL and concomitant SLE and APS.
Background:Germline BRCA1 (gBRCA1) pathogenic variants (PVs) are the most common cause of inherited breast cancer. Preclinical studies have shown that homozygous loss of BRCA1 in T cells impairs T cell-mediated antitumour immunity. However, the consequences of heterozygous gBRCA1 on human T cell function are unknown. Patients and methods:We collected peripheral blood mononuclear cells (PBMCs) from healthy donors and treatment-naïve patients diagnosed with early-stage breast cancer with or without a gBRCA1 PV. T cells derived from PBMC were activated in vitro and analysed for phenotypic and functional differences. To investigate the impact of gBRCA1 PV on tumour-infiltrating lymphocytes (TILs), we carried out T cell receptor (TCR) sequencing and whole exome sequencing on triple negative breast cancer (TNBC) tumours from gBRCA1 carriers and wild-type (WT) patients. Results:Patients with gBRCA1 PV had significantly reduced circulating T cell counts compared with WT individuals. BRCA1 protein was highly expressed in healthy donor activated T cells, but significantly reduced expression was observed in gBRCA1 patients compared with WT. Activated T cells from gBRCA1 carriers displayed a distinct transcriptional profile with significantly impaired proliferation and decreased effector molecule production. TCR sequencing of TILs extracted from TNBC tumours from gBRCA1 carriers revealed significantly more hyperexpanded T cell clones than in WT patients, with hyperexpansion correlating with increased tumour mutation burden (R 2 = 0.86), particularly frameshift mutations (R 2 = 0.72). Conclusions:Circulating T cells from gBRCA1 carriers exhibit intrinsic T cell dysfunction. This phenotype may be clinically relevant in the context of antitumour immunity, cancer development, progression and response to immunotherapy treatment.
Background:Brown adipose tissue (BAT) activity has been suggested to play a role in cancer progression. Previous studies have shown that BAT activity is higher in patients with cancer, and that BAT volume is a predictor of tumour recurrence and mortality in patients with cancer, but the data on melanoma are limited. Patients and methods:Here, we re-analysed 2-fluoro-2-deoxy-D-glucose positron emission tomography-computed tomography (FDG-PET-CT) images from 135 patients with cutaneous melanoma treated at Turku University Hospital between 2012 and 2021 to assess associations among BAT, melanoma progression, patient survival, and patient weight. We applied a three-stage universal BAT threshold definition using predetermined standardised uptake value thresholds of 0.8, 1.0, and 1.2 g/ml, given the retrospective nature of our study. Of the 135 patients (81 men and 54 women; median age 61 years, interquartile range 54-71), 40 (29.6%), 24 (17.8%), and 19 (14.1%) were BAT-positive at the 0.8, 1.0, and 1.2 g/ml thresholds, respectively. Results:Our results showed that patients with active melanoma on FDG-PET-CT imaging were more frequently BAT-positive at the 0.8 threshold (P = 0.026) and 1.0 threshold (P = 0.016). We also found a significantly higher BAT volume among patients who survived the observation period (0.8, 1.0, and 1.2 g/ml thresholds; P = 0.018, P = 0.038, and P = 0.571, respectively) and those who did not relapse (0.8, 1.0, and 1.2 g/ml thresholds; P = 0.631, P = 0.012, and P = 0.030, respectively). Conclusions:No association between BAT positivity and relapse-free survival or overall survival was observed at any threshold. Although higher BAT volumes were observed in subgroups of patients who survived or did not relapse, these findings were not supported by survival analyses and should be considered exploratory.
Background:Immune checkpoint inhibitor (ICI) therapy improves survival in head and neck squamous cell carcinoma (HNSCC), yet only some patients benefit, highlighting the need for early identification of responders and functional evaluation of tumor-immune interactions. We developed an autologous coculture platform combining patient-derived organoids (PDOs) with peripheral blood mononuclear cells (PBMCs) to assess immune responses and correlate preclinical findings with clinical outcomes. Material and methods:PDOs from HNSCC specimens were expanded and cocultured with PBMCs, followed by treatment with ICIs targeting programmed cell death protein 1 (PD-1), lymphocyte-activation gene 3 (LAG-3), or their combination. Results:PBMCs were enriched for CD4+ and CD8+ T cells (∼90%) and CD16+CD56+ natural killer cells, with baseline exhaustion (LAG-3+, CTLA-4+). T cell-mediated tumor killing occurred in 44% of cases and was accompanied by Th1/Th17 cytokine signatures. ICI treatment modestly shifted T-cell phenotypes, reducing exhaustion (PD-1) and enriching cytolytic subsets (CD4+IFN-γ+, CD8+TOX+). Interleukin (IL)-17A emerged as a relevant cytokine, correlating with FasL secretion and suggesting activation of the Fas/FasL axis. High circulating IL-17A levels in treatment-naïve HNSCC patients were associated with improved survival, whereas other cytokines showed no prognostic value. In a noninterventional observational trial, increases in IL-17A positively correlated with favorable ICI responses, indicating its potential as a biomarker. PDO viability decreased in response to ICIs in a patient-specific manner, consistent with cytokine signatures and clinical outcomes. Conclusion:Collectively, this autologous PDO-PBMC coculture platform, combined with longitudinal cytokine profiling, captures patient-specific tumor-immune interactions and interpatient heterogeneity, providing a framework for real-time response prediction, biomarker discovery, and personalized immunotherapy in HNSCC.
Background:Immune checkpoint inhibition (ICI) provides durable benefit across several cancers but carries a substantial risk of immune-related adverse events (irAEs). Management relies primarily on corticosteroids, yet responses are variable and difficult to predict. In this observational study, we explored whether early immune phenotypes in fresh whole blood reflect organ-specific irAEs and corticosteroid responsiveness. Materials and methods:We prospectively enrolled patients receiving ICI who developed grade 2-4 colitis, hepatitis, or nephritis. Peripheral blood was collected at irAE onset (T0) and during early management (T1). Immune profiling was carried out on fresh blood by flow cytometry and unsupervised clustering. Results:Twenty-seven patients were included (colitis n = 13, hepatitis n = 11, nephritis n = 3). Organ-specific immune patterns were observed at irAE onset: hepatitis showed higher CD27+ T cells and naive B cells; nephritis exhibited reduced T cells and expansion of CD16- natural killer cells; and colitis displayed a more variable profile. Patients were categorized as corticosteroid responders (n = 8), refractory (n = 9), or relapse (n = 10). At T0, responders showed more CD38+ T cells and naive B cells with fewer classical monocytes, relapse patients had increased memory B cells, and refractory patients had lower CD27+ T cells and elevated classical monocytes. Immune shifts from T0 to T1 were modest, though fold-change analyses revealed response-associated patterns, such as preserved CD56+ T cells and monocyte expansion in responders. Conclusions:In this explorative study, fresh-blood immunophenotyping suggested associations between irAE type, early immune signatures, and corticosteroid response. These hypothesis-generating and preliminary findings support further research into a more individualized immune-guided approach to irAE management.
Background: Myxofibrosarcoma (MFS) is a subtype of soft-tissue sarcoma for which local treatment includes neoadjuvant radiotherapy (nRT) followed by surgery. The impact of nRT on the MFS tumor immune microenvironment remains unexplored. Materials and methods: Paired pre-nRT biopsy and post-nRT surgery samples from 31 MFS patients were retrospectively collected. The intratumoral density of T cells, cytotoxic T cells, regulatory T cells, helper T cells, B cells and natural killer (NK) cells and expression of programmed death-ligand 1 were quantified using multiplex immunohistochemistry (mIHC). mIHC marker densities were compared between pre- and post-nRT samples, and their associations with clinicopathological characteristics and patient outcomes were assessed. Results: There was substantial interpatient heterogeneity in mIHC marker densities, both pre- and post-nRT. A significant reduction in cytotoxic T cell {205 [interquartile range (IQR) 674] pre-nRT versus 58 (IQR 205) post-nRT, P = 0.030} and helper T cell [220 (IQR 343) pre-nRT versus 74 (IQR 110) post-nRT, P = 0.011] densities, alongside an increase in NK cell [2 (IQR 8) pre-nRT versus 7 (IQR 16) post-nRT, P = 0.050] infiltration, was observed following nRT. There were no statistically significant associations between mIHC marker densities measured before or after nRT, or their changes over time, and patient outcomes. Conclusions: Conventional nRT is associated with reduced cytotoxic and helper T cell infiltration and increased presence of NK cells in MFS. These results suggest that conventional nRT reduces inflammatory aspects of MFS tumors.
Background: Acute myeloid leukemia (AML) is a high-risk malignancy, particularly in patients with primary induction failure or relapsed/refractory disease. CD117 (c-Kit), expressed on both leukemic blasts and normal hematopoietic stem and progenitor cells (HSPCs), represents a potential therapeutic target but poses challenges due to the risk of severe myelotoxicity. Materials and methods: Retrospective flow cytometry analyses of samples from 27 AML patients and AML cell lines were carried out to assess CD117 expression. Second-generation CD117-specific chimeric antigen receptor (CAR) T cells were generated by either retroviral transduction or in vitro-transcribed (IVT) messenger RNA (mRNA) electroporation. The mRNA-based CD117 CAR T cells were evaluated for viability, immunophenotype, cytotoxic activity, and toxicity toward primary HSPCs using clonogenic assays, and compared with retroviral-based counterparts. Results: CD117 was expressed in AML patient samples and cell lines at varying levels. CD117 CAR T cells demonstrated potent and specific cytotoxicity against AML cells. The mRNA-based CAR T cells exhibited high transfection efficiency, good viability, and an immunophenotype similar to non-transduced T cells, and were functionally competent as early as 2 h post-electroporation. In long-term co-culture with a high tumor burden, repeated dosing of mRNA CAR T cells effectively eliminated CD117+ cells, comparable to viral vector-based CAR T cells. Notably, residual mRNA CAR T cells following AML clearance showed no detectable CAR expression and preserved HSPC colony-forming capacity. Conclusions: Our in vitro studies suggest the potential use of mRNA CD117 CAR T cells as a non-genotoxic preconditioning strategy for patients with high-risk or refractory AML.
Background:LOAd703 is a tumor microenvironment (TME) gene-engineering adenovirus encoding the immunostimulatory transgenes trimerized membrane-bound CD40 ligand (CD40L) and 4-1BB ligand (4-1BBL). Upon administration in the TME, the transgenes are expressed in various cell types to engage both the tumor and its stroma to activate antitumor immunity. CD40-CD40L interaction causes dendritic cell maturation and stimulates T helper 1-type immune responses, whereas 4-1BB-4-1BBL signaling protects T cells and natural killer cells from activation-induced cell death and promotes lymphocyte proliferation. LOAd703 replication with subsequent oncolysis is restricted to cancer cells. Patients and methods:In the dose-escalating part of this clinical study (NCT03225989), LOAd703 was increased according to a standard 3 + 3 design in patients with advanced solid malignancies. LOAd703 was administered every 2 weeks by ultrasound-guided intratumoral injections, combined with a standard-of-care or immune-conditioning gemcitabine-based chemotherapy regimen. The primary endpoint was tolerability. Results:Three dose levels of LOAd703 were evaluated in 10 patients. Treatment was overall safe and well tolerated. The most common side-effects assessed as secondary to LOAd703 were pyrexia, fatigue and headache. All LOAd703-attributed adverse events were of grade 1-2, and the majority were transient and emerged shortly after administration. One patient developed cytokine release syndrome grade 2. The maximum tolerated dose was not reached. Median overall survival was 8.4 months, and the overall response rate was 20%. A trend of higher interferon-gamma (IFN-γ) plasma levels in the highest LOAd703 dose cohort was observed. Conclusion:The acceptable toxicity associated with LOAd703 and chemotherapy, combined with signs of clinical benefit in poor prognostic cancer patients, warrant further studies.
Chimeric antigen receptor (CAR) cellular therapy, particularly CAR-T cells, has revolutionized the treatment of hematologic malignancies. However, these therapies show limited efficacy against solid tumors, in part due to the inefficient trafficking of effector cells to the tumor. This review explores the potential of engineering natural and synthetic G protein-coupled receptors (GPCRs) to overcome this migratory hurdle. Chemokine receptors have been the most used GPCR family in this setting. Engineering effector immune cells to express chemokine receptors that match tumor-derived chemokines has been shown to increase their chemotaxis and to improve antitumor efficacy in preclinical models. In addition to improved migration, chemokine receptor engineering can also have additional benefits, such as remodeling of the tumor microenvironment and metabolic rewiring of engineered cells. However, the effectiveness of this approach is limited by the tumor-specific and heterogeneous chemokine milieu. Emerging strategies make use of synthetic GPCRs and could overcome some of these limitations using chemogenetic and optogenetic approaches. Here, mutated GPCRs binding only to specific and orthogonal ligands or light-sensitive channels are used for cell modulation and trafficking. Equipping cells with these synthetic GPCRs allows for precise and stimulus-controlled immune cell migration. Together, natural and synthetic GPCR engineering form promising approaches to enhance immune cell trafficking, persistence, and efficacy.
Background:Inflammatory plasma proteins have been associated with poorer clinical outcomes in melanoma patients treated with immune checkpoint inhibitors (ICIs). Furthermore, the plasma levels of thymidine kinase activity (TKa), an indicator of proliferation, have also been associated with adverse outcomes in several cancers. The aim of this study was to explore the relationship between TKa, systemic inflammation and survival in ICI-treated melanoma patients. Materials and methods:Fifty-eight patients with unresectable metastatic melanoma who received anti-programmed cell death protein 1 monotherapy were included in the study. Pretreatment plasma samples were analyzed for TKa levels (DiviTum® TKa assay; Biovica) and inflammatory proteins using the Olink® Target 96 Inflammation panel, and these biomarkers were correlated with patient survival. Results:A panel of five unfavorable biomarkers was selected, comprising TKa, C-X-C motif chemokine ligand 8, C-C motif chemokine ligand 3, hepatocyte growth factor and S100 calcium-binding protein A12. An increasing number of elevated biomarkers in the panel was associated with a progressively worse prognosis. In the univariate analysis, when patients with four to five unfavorable biomarkers were compared with those with none, the hazard ratio (HR) was 2.43 [95% confidence interval (CI) 0.91-6.44, P = 0.075] for progression-free survival (PFS) and 10.62 (95% CI 3.11-36.26, P < 0.001) for overall survival (OS). When adjusting for sex, age, stage and lactate dehydrogenase (LDH), the HR was 1.97 (95% CI 0.61-6.41, P = 0.26) for PFS and 11.03 (95% CI 2.53-48.12, P = 0.001) for OS. Conclusion:This multi-biomarker panel was independently associated with poor OS in ICI-treated melanoma patients. This finding suggests a prognostic value of the panel, indicating its potential relevance for stratifying patients by risk, although further validation in larger cohorts is warranted.
