Publikasjoner
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Fjørtoft, Marit Otterlei; Garred, Øystein; Lingjærde, Ole Christian; Johannesen, Karin Teien; Ottestad, Lars & Bergheim, Inger Riise
[Vis alle 29 forfattere av denne artikkelen]
(2025).
Single-cell analysis reveals tumour size as a key driver of immune cell profile alterations in primary breast tumours and corresponding lymph nodes.
BJC Reports.
3(1).
doi:
10.1038/s44276-025-00152-3.
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At diagnosis, 30–40% of women with breast cancer have metastases in sentinel (SN) or axillary lymph nodes (ALN). Nodal status is a strong prognostic factor and guides treatment decisions. Immune checkpoint inhibition has shown some efficacy, which can increase in the neoadjuvant setting. A better understanding of how tumour cells in primary tumours and metastatic lymph nodes shape the local immune microenvironment may provide clues for more individualized therapeutic interventions.
Methods
We conducted deep immunophenotypic analysis of 29 primary breast tumours and 36 lymph nodes from 38 patients with primary operable breast cancer.
Results
The immune profile of the primary tumour was not predictive of the lymph node immune profile or metastatic status. Primary tumours showed prominent CD8 T cell exhaustion and activated regulatory T cells, and the frequencies of these subsets were associated with tumour size. The immune cell profile in lymph nodes were different from the profile in primary tumours, except for the ALN+ nodes, which displayed a T-cell profile more similar to primary tumours. The frequencies of the T cell subsets in lymph nodeswere associated with metastatic size. Tumour cells from smaller metastases exhibited a distinct phenotype compared to those from larger tumour deposits, and the size of the tumour cell deposit impacted the local immune cell composition.
Conclusion
The tumour size of primary tumours and metastatic size in lymph nodes are the main drivers of changes in immune cell composition
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Otterlei, Marit; Huse, Kanutte & Rye, Inga Hansine
(2024).
The Tumor Immune Microenvironment in Breast Cancer Progression.
Acta Oncologica.
ISSN 0284-186X.
63,
s. 359–367.
doi:
10.2340/1651-226X.2024.33008.
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Background: The tumor microenvironment significantly influences breast cancer development, progression, and metastasis. Various immune cell populations, including T cells, B cells, NK cells, and myeloid cells exhibit diverse functions in different breast cancer subtypes, contributing to both anti-tumor and pro-tumor activities. Purpose: This review provides an overview of the predominant immune cell populations in breast cancer subtypes, elucidating their suppressive and prognostic effects. We aim to outline the role of the immune microenvironment from normal breast tissue to invasive cancer and distant metastasis. Methods: A comprehensive literature review was conducted to analyze the involvement of immune cells throughout breast cancer progression. Results: In breast cancer, tumors exhibit increased immune cell infiltration compared to normal tissue. Variations exist across subtypes, with higher levels observed in triple-negative and HER2+ tumors are linked to better survival. In contrast, ER+ tumors display lower immune infiltration, associated with poorer outcomes. Furthermore, metastatic sites commonly exhibit a more immunosuppressive microenvironment. Conclusion: Understanding the complex interaction between tumor and immune cells during breast cancer progression is essential for future research and the development of immune-based strategies. This comprehensive understanding may pave the way for more effective treatment approaches and improved patients outcomes.
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Chauhan, Sudhir Kumar; Dunn, Claire; Andresen, Nikolai Kragøe; Røssevold, Andreas Hagen; Skorstad, Gjertrud & Sike, Adam
[Vis alle 11 forfattere av denne artikkelen]
(2024).
Peripheral immune cells in metastatic breast cancer patients display a systemic immunosuppressed signature consistent with chronic inflammation.
NPJ breast cancer.
10(1).
doi:
10.1038/s41523-024-00638-2.
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Immunotherapies blocking the PD-1/PD-L1 checkpoint show some efficacy in metastatic breast cancer (mBC) but are often hindered by immunosuppressive mechanisms. Understanding these mechanisms is crucial for personalized treatments, with peripheral blood monitoring representing a practical alternative to repeated biopsies. In the present study, we performed a comprehensive mass cytometry analysis of peripheral blood immune cells in 104 patients with HER2 negative mBC and 20 healthy donors (HD). We found that mBC patients had significantly elevated monocyte levels and reduced levels of CD4+ T cells and plasmacytoid dendritic cells, when compared to HD. Furthermore, mBC patients had more effector T cells and regulatory T cells, increased expression of immune checkpoints and other activation/exhaustion markers, and a shift to a Th2/Th17 phenotype. Furthermore, T-cell phenotypes identified by mass cytometry correlated with functionality as assessed by IFN-γ production. Additional analysis indicated that previous chemotherapy and CDK4/6 inhibition impacted the numbers and phenotype of immune cells. From 63 of the patients, fresh tumor samples were analyzed by flow cytometry. Paired PBMC-tumor analysis showed moderate correlations between peripheral CD4+ T and NK cells with their counterparts in tumors. Further, a CD4+ T cell cluster in PBMCs, that co-expressed multiple checkpoint receptors, was negatively associated with CD4+ T cell tumor infiltration. In conclusion, the identified systemic immune signatures indicate an immune-suppressed environment in mBC patients who had progressed/relapsed on standard treatments and is consistent with ongoing chronic inflammation. These activated immuno-suppressive mechanisms may be investigated as therapeutic targets, and for use as biomarkers of response or treatment resistance.
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Meriranta, Leo; Sorri, Selma; Huse, Kanutte; Liu, Xiaonan; Spasevska, Ivana & Safar, Zadia
[Vis alle 15 forfattere av denne artikkelen]
(2024).
Disruption of KLHL6 Fuels Oncogenic Antigen Receptor Signaling in B-Cell Lymphoma.
Blood Cancer Discovery.
ISSN 2643-3230.
5(5),
s. 331–352.
doi:
10.1158/2643-3230.BCD-23-0182.
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Pathomechanisms that activate oncogenic B-cell receptor (BCR) signaling in diffuse large B-cell lymphoma (DLBCL) are largely unknown. Kelch-like family member 6 (KLHL6) encoding a substrate-adapter for Cullin-3-RING E3 ubiquitin ligase with poorly established targets is recurrently mutated in DLBCL. By applying high-throughput protein interactome screens and functional characterization, we discovered that KLHL6 regulates BCR by targeting its signaling subunits CD79A and CD79B. Loss of physiologic KLHL6 expression pattern was frequent among the MCD/C5-like activated B-cell DLBCLs and was associated with higher CD79B levels and dismal outcome. Mutations in the bric-a-brac tramtrack broad domain of KLHL6 disrupted its localization and heterodimerization and increased surface BCR levels and signaling, whereas Kelch domain mutants had the opposite effect. Malfunctions of KLHL6 mutants extended beyond proximal BCR signaling with distinct phenotypes from KLHL6 silencing. Collectively, our findings uncover how recurrent mutations in KLHL6 alter BCR signaling and induce actionable phenotypic characteristics in DLBCL. Significance: Oncogenic BCR signaling sustains DLBCL cells. We discovered that Cullin-3-RING E3 ubiquitin ligase substrate-adapter KLHL6 targets BCR heterodimer (CD79A/CD79B) for ubiquitin-mediated degradation. Recurrent somatic mutations in the KLHL6 gene cause corrupt BCR signaling by disrupting surface BCR homeostasis. Loss of KLHL6 expression and mutant-induced phenotypes associate with targetable disease characteristics in B-cell lymphoma. See related commentary by Leveille et al. See related commentary by Corcoran et al.
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Spasevska, Ivana; Sharma, Ankush; Steen, Chloe Beate; Josefsson, Sarah Elisabet; Blaker, Yngvild Nuvin & Kolstad, Arne
[Vis alle 26 forfattere av denne artikkelen]
(2023).
Diversity of intratumoral regulatory T cells in B-cell non-Hodgkin lymphoma.
Blood Advances.
ISSN 2473-9529.
7(23),
s. 7216–7230.
doi:
10.1182/bloodadvances.2023010158.
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Abstract Tumor-infiltrating regulatory T cells (Tregs) contribute to an immunosuppressive tumor microenvironment. Despite extensive studies, the prognostic impact of tumor-infiltrating Tregs in B-cell non-Hodgkin lymphomas (B-NHLs) remains unclear. Emerging studies suggest substantial heterogeneity in the phenotypes and suppressive capacities of Tregs, emphasizing the importance of understanding Treg diversity and the need for additional markers to identify highly suppressive Tregs. Here, we applied single-cell RNA sequencing and T-cell receptor sequencing combined with high-dimensional cytometry to decipher the heterogeneity of intratumoral Tregs in diffuse large B-cell lymphoma and follicular lymphoma (FL), compared with that in nonmalignant tonsillar tissue. We identified 3 distinct transcriptional states of Tregs: resting, activated, and unconventional LAG3+FOXP3− Tregs. Activated Tregs were enriched in B-NHL tumors, coexpressed several checkpoint receptors, and had stronger immunosuppressive activity compared with resting Tregs. In FL, activated Tregs were found in closer proximity to CD4+ and CD8+ T cells than other cell types. Furthermore, we used a computational approach to develop unique gene signature matrices, which were used to enumerate each Treg subset in cohorts with bulk gene expression data. In 2 independent FL cohorts, activated Tregs was the major subset, and high abundance was associated with adverse outcome. This study demonstrates that Tregs infiltrating B-NHL tumors are transcriptionally and functionally diverse. Highly immunosuppressive activated Tregs were enriched in tumor tissue but absent in the peripheral blood. Our data suggest that a deeper understanding of Treg heterogeneity in B-NHL could open new paths for rational drug design, facilitating selective targeting to improve antitumor immunity.
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Huse, Kanutte; Bai, Baoyan; Hilden, Vera Irene; Bollum, Lise Kristin; Våtsveen, Thea Kristin & Munthe, Ludvig Andre
[Vis alle 10 forfattere av denne artikkelen]
(2022).
Mechanism of CD79A and CD79B support for IgM+ B cell fitness through B cell receptor surface expression.
Journal of Immunology.
ISSN 0022-1767.
209(10),
s. 2042–2053.
doi:
10.4049/jimmunol.2200144.
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he BCR consists of surface-bound Ig and a heterodimeric signaling unit comprised of CD79A and CD79B. Upon cognate Ag recognition, the receptor initiates important signals for B cell development and function. The receptor also conveys Ag-independent survival signals termed tonic signaling. Although the requirement of a CD79A/CD79B heterodimer for BCR complex assembly and surface expression is well established based on mice models, few studies have investigated this in human mature B cells. In this study, we found that human tonsillar B cells with high surface expression of IgM or IgG had potentiated BCR signaling compared with BCRlow cells, and high IgM expression in germinal center B cells was associated with reduced apoptosis. We explored the mechanism for IgM surface expression by CRISPR/Cas9-induced deletion of CD79A or CD79B in four B lymphoma cell lines. Deletion of either CD79 protein caused loss of surface IgM in all cell lines and reduced fitness in three. From two cell lines, we generated stable CD79A or CD79B knockout clones and demonstrated that loss of CD79A or CD79B caused a block in N-glycan maturation and accumulation of immature proteins, compatible with retention of BCR components in the endoplasmic reticulum. Rescue experiments with CD79B wild-type restored surface expression of CD79A and IgM with mature glycosylation, whereas a naturally occurring CD79B G137S mutant disrupting CD79A/CD79B heterodimerization did not. Our study highlights that CD79A and CD79B are required for surface IgM expression in human B cells and illuminates the importance of the IgM expression level for signaling and fitness.
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Rye, Inga Hansine; Huse, Kanutte; Josefsson, Sarah Elisabet; Kildal, Wanja; Danielsen, Håvard Emil Greger & Schlichting, Ellen
[Vis alle 28 forfattere av denne artikkelen]
(2021).
Breast cancer metastasis: immune profiling of lymph nodes reveals exhaustion of effector T cells and immunosuppression.
Molecular Oncology.
ISSN 1574-7891.
16(1),
s. 88–103.
doi:
10.1002/1878-0261.13047.
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Sentinel lymph nodes are the first nodes draining the lymph from a breast and could reveal early changes in the host immune system upon dissemination of breast cancer cells. To investigate this, we performed single-cell immune profiling of lymph nodes with and without metastatic cells. Whereas no significant changes were observed for B-cell and natural killer (NK)-cell subsets, metastatic lymph nodes had a significantly increased frequency of CD8 T cells and a skewing toward an effector/memory phenotype of CD4 and CD8 T cells, suggesting an ongoing immune response. Additionally, metastatic lymph nodes had an increased frequency of TIGIT (T-cell immunoreceptor with Ig and ITIM domains)-positive T cells with suppressed TCR signaling compared with non-metastatic nodes, indicating exhaustion of effector T cells, and an increased frequency of regulatory T cells (Tregs) with an activated phenotype. T-cell alterations correlated with the percentage of metastatic tumor cells, reflecting the presence of metastatic tumor cells driving T effector cells toward exhaustion and promoting immunosuppression by recruitment or increased differentiation toward Tregs. These results show that immune suppression occurs already in early stages of tumor progression.
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Trifonas, Idun D. Rein; Notø, Heidi Ødegaard; Bostad, Monica; Huse, Kanutte & Stokke, Trond
(2020).
Cell Cycle Analysis and Relevance for Single‐Cell Gating in Mass Cytometry.
Cytometry Part A.
ISSN 1552-4922.
97(8),
s. 832–844.
doi:
10.1002/cyto.a.23960.
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Cell cycle analysis by mass cytometry (MC) is hampered by the poor resolution of the Iridium‐labeled DNA intercalator compared to DNA‐specific fluorescent dyes. We report here a minimum cell cycle panel for MC consisting of Ir‐intercalator (DNA content), IdU (S phase), anti‐pS28HistoneH3 (mitosis), anti‐CDT1 (G1 phase) and anti‐Geminin (non‐G1 phases). Cell cycle distributions obtained by MC were not significantly different from fluorescence flow cytometry results (r2 = 0.98, P < 0.001). Further subdivision of the G1 and G2 phases could be done with anti‐pS780RB1 (late G1) and anti‐PLK1 (late G2), respectively. A disadvantage of MC is that aggregates of cells cannot easily be removed while retaining all single cells. We have developed an analysis pipeline including unsupervised clustering by FlowSOM and subsequent single‐cell gating. When performed on cells stained with the cell cycle panel, this analysis pipeline successfully identified debris, dead/apoptotic cells, nonsingle‐cell populations and the major cell cycle phases. The presented cell cycle panel and analysis pipeline thus achieves true single‐cell analysis at the same time as any additional channels in the panel are open for phenotyping and cell cycle‐resolved expression or modification analysis. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry.
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Sonnet, Friederike; Namork, Ellen; Stylianou, Eva; Gaare-Olstad, Ingvild; Huse, Kanutte & Andorf, Sandra
[Vis alle 9 forfattere av denne artikkelen]
(2020).
Reduced polyfunctional T cells and increased cellular activation markers in adult allergy patients reporting adverse reactions to food.
BMC Immunology.
21,
s. 1–14.
doi:
10.1186/s12865-020-00373-w.
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Background The underlying cellular mechanisms causing adverse reactions to food are complex and still not fully understood. Therefore, in this study we aimed to identify functional and/or phenotypical immune cell signatures characteristic for adult patients reporting adverse reactions to food. By mass cytometry, we performed high-dimensional profiling of peripheral blood mononuclear cells (PBMC) from adult patients reporting adverse reactions to food and healthy controls. The patients were grouped according to sIgE-positive or sIgE-negative serology to common food and inhalant allergens. Two broad antibody panels were used, allowing determination of major immune cell populations in PBMC, as well as activation status, proliferation status, and cytokine expression patterns after PMA/ionomycin-stimulation on a single cell level. Results By use of data-driven algorithms, several cell populations were identified showing significantly different marker expression between the groups. Most striking was an impaired frequency and function of polyfunctional CD4+ and CD8+ T cells in patients reporting adverse reactions to food compared to the controls. Further, subpopulations of monocytes, T cells, and B cells had increased expression of functional markers such as CD371, CD69, CD25, CD28, and/or HLA-DR as well as decreased expression of CD23 in the patients. Most of the differing cell subpopulations were similarly altered in the two subgroups of patients. Conclusion Our results suggest common immune cell features for both patient subgroups reporting adverse reactions to food, and provide a basis for further studies on mechanistic and diagnostic biomarker studies in food allergy.
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Josefsson, Sarah Elisabet; Beiske, Klaus; Blaker, Yngvild Nuvin; Førsund, Mette S; Holte, Harald & Østenstad, Bjørn
[Vis alle 15 forfattere av denne artikkelen]
(2019).
TIGIT and PD-1 mark intratumoral T cells with reduced effector function in B-cell non-Hodgkin lymphoma.
Cancer immunology research.
ISSN 2326-6066.
7(3),
s. 355–362.
doi:
10.1158/2326-6066.CIR-18-0351.
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Checkpoint blockade can reverse T-cell exhaustion and promote antitumor responses. Although blocking the PD-1 pathway has been successful in Hodgkin lymphoma, response rates have been modest in B-cell non-Hodgkin lymphoma (NHL). Coblockade of checkpoint receptors may therefore be necessary to optimize antitumor T-cell responses. Here, characterization of coinhibitory receptor expression in intratumoral T cells from different NHL types identified TIGIT and PD-1 as frequently expressed coinhibitory receptors. Tumors from NHL patients were enriched in CD8+ and CD4+ T effector memory cells that displayed high coexpression of TIGIT and PD-1, and coexpression of these checkpoint receptors identified T cells with reduced production of IFNγ, TNFα, and IL2. The suppressed cytokine production could be improved upon in vitro culture in the absence of ligands. Whereas PD-L1 was expressed by macrophages, the TIGIT ligands CD155 and CD112 were expressed by lymphoma cells in 39% and 50% of DLBCL cases and in some mantle cell lymphoma cases, as well as by endothelium and follicular dendritic cells in all NHLs investigated. Collectively, our results show that TIGIT and PD-1 mark dysfunctional T cells and suggest that TIGIT and PD-1 coblockade should be further explored to elicit potent antitumor responses in patients with NHL.
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Köksal, Hakan; Dillard, Pierre; Josefsson, Sarah Elisabet; Maggadottir, Solrun Melkorka; Pollmann, Sylvie Mireille & Fåne, Anne
[Vis alle 16 forfattere av denne artikkelen]
(2019).
Preclinical development of CD37CAR T-cell therapy for treatment of B-cell lymphoma.
Blood Advances.
ISSN 2473-9529.
3(8),
s. 1230–1243.
doi:
10.1182/bloodadvances.2018029678.
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T cells modified to express chimeric antigen receptor (CAR) targeting CD19 (CD19CAR) have produced remarkable clinical responses in patients with relapsed/refractory B-cell acute lymphoblastic leukemia. CD19CAR T-cell therapy has also demonstrated prominent effects in B-cell non-Hodgkin lymphoma (B-NHL) patients. However, a subset of patients who relapse after CD19CAR T-cell therapy have outgrowth of CD19− tumor cells. Hence, development of alternative CARs targeting other B-cell markers represents an unmet medical need for B-cell acute lymphoblastic leukemia and B-NHL. Here, we confirmed previous data by showing that, overall, B-NHL has high expression of CD37. A second-generation CD37CAR was designed, and its efficacy in T cells was compared with that of CD19CAR. In vitro assessment of cytotoxicity and T-cell function upon coculture of the CAR T cells with different target B-cell lymphoma cell lines demonstrated comparable efficacy between the 2 CARs. In an aggressive B-cell lymphoma xenograft model, CD37CAR T cells were as potent as CD19CAR T cells in controlling tumor growth. In a second xenograft model, using U2932 lymphoma cells containing a CD19− subpopulation, CD37CAR T cells efficiently controlled tumor growth and prolonged survival, whereas CD19CAR T cells had limited effect. We further show that, unlike CD19CAR, CD37CAR was not sensitive to antigen masking. Finally, CD37CAR reactivity was restricted to B-lineage cells. Collectively, our results demonstrated that CD37CAR T cells also can effectively eradicate B-cell lymphoma tumors when CD19 antigen expression is lost and support further clinical testing for patients with relapsed/refractory B-NHL.
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Josefsson, Sarah Elisabet Göthberg; Huse, Kanutte; Kolstad, Arne; Beiske, Klaus; Pende, Daniela & Steen, Chloe Beate
[Vis alle 13 forfattere av denne artikkelen]
(2017).
T cells expressing checkpoint receptor TIGIT are enriched in follicular lymphoma tumors and characterized by reversible suppression of T-cell receptor signaling.
Clinical Cancer Research.
ISSN 1078-0432.
24(4),
s. 870–881.
doi:
10.1158/1078-0432.CCR-17-2337.
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Purpose: T cells infiltrating follicular lymphoma (FL) tumors are considered dysfunctional, yet the optimal target for immune checkpoint blockade is unknown. Characterizing coinhibitory receptor expression patterns and signaling responses in FL T-cell subsets might reveal new therapeutic targets. Experimental Design: Surface expression of 9 coinhibitory receptors governing T-cell function was characterized in T-cell subsets from FL lymph node tumors and from healthy donor tonsils and peripheral blood samples, using high-dimensional flow cytometry. The results were integrated with T-cell receptor (TCR)-induced signaling and cytokine production. Expression of T-cell immunoglobulin and ITIM domain (TIGIT) ligands was detected by immunohistochemistry. Results: TIGIT was a frequently expressed coinhibitory receptor in FL, expressed by the majority of CD8 T effector memory cells, which commonly coexpressed exhaustion markers such as PD-1 and CD244. CD8 FL T cells demonstrated highly reduced TCR-induced phosphorylation (p) of ERK and reduced production of IFNγ, while TCR proximal signaling (p-CD3ζ, p-SLP76) was not affected. The TIGIT ligands CD112 and CD155 were expressed by follicular dendritic cells in the tumor microenvironment. Dysfunctional TCR signaling correlated with TIGIT expression in FL CD8 T cells and could be fully restored upon in vitro culture. The costimulatory receptor CD226 was downregulated in TIGIT+ compared with TIGIT− CD8 FL T cells, further skewing the balance toward immunosuppression. Conclusions: TIGIT blockade is a relevant strategy for improved immunotherapy in FL. A deeper understanding of the interplay between coinhibitory receptors and key T-cell signaling events can further assist in engineering immunotherapeutic regimens to improve clinical outcomes of cancer patients.
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Bollum, Lise Kristin; Huse, Kanutte; Oksvold, Morten Pedersen; Bai, Baoyan; Hilden, Vera Irene & Forfang, Lise
[Vis alle 10 forfattere av denne artikkelen]
(2017).
BMP-7 induces apoptosis in human germinal center B cells and is influenced by TGF-β receptor type I ALK5.
PLOS ONE.
12(5).
doi:
10.1371/journal.pone.0177188.
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Selection and maturation of B cells into plasma cells producing high-affinity antibodies occur in germinal centers (GC). GCs form transiently in secondary lymphoid organs upon antigen challenge, and the GC reaction is a highly regulated process. TGF-β is a potent negative regulator, but the influence of other family members including bone morphogenetic proteins (BMPs) is less known. Studies of human peripheral blood B lymphocytes showed that BMP-6 suppressed plasmablast differentiation, whereas BMP-7 induced apoptosis. Here, we show that human naïve and GC B cells had a strikingly different receptor expression pattern. GC B cells expressed high levels of BMP type I receptor but low levels of type II receptors, whereas naïve B cells had the opposite pattern. Furthermore, GC B cells had elevated levels of downstream signaling components SMAD1 and SMAD5, but reduced levels of the inhibitory SMAD7. Functional assays of GC B cells revealed that BMP-7 suppressed the viability-promoting effect of CD40L and IL-21, but had no effect on CD40L- and IL-21-induced differentiation into plasmablasts. BMP-7-induced apoptosis was counteracted by a selective TGF-β type I receptor (ALK4/5/7) inhibitor, but not by a selective BMP receptor type I inhibitor. Furthermore, overexpression of truncated ALK5 in a B-cell line counteracted BMP-7-induced apoptosis, whereas overexpression of truncated ALK4 had no effect. BMP-7 mRNA and protein was readily detected in tonsillar B cells, indicating a physiological relevance of the study. Altogether, we identified BMP-7 as a negative regulator of GC B-cell survival. The effect was counteracted by truncated ALK5, suggesting greater complexity in regulating BMP-7 signaling than previously believed.
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Wogsland, Cara Ellen; Greenplate, Allison Rae; Kolstad, Arne; Myklebust, June; Irish, Jonathan Michael & Huse, Kanutte
(2017).
Mass cytometry of follicular lymphoma tumors reveals intrinsic heterogeneity in proteins including HLA-DR and a deficit in nonmalignant plasmablast and germinal center B-cell populations.
Cytometry Part B: Clinical Cytometry.
ISSN 1552-4949.
92(1),
s. 79–87.
doi:
10.1002/cyto.b.21498.
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Polikowsky, Hannah G.; Wogsland, Cara E.; Diggins, Kirsten E.; Huse, Kanutte & Irish, Jonathan M.
(2015).
Cutting edge: Redox signaling hypersensitivity distinguishes human germinal center B cells.
Journal of Immunology.
ISSN 0022-1767.
195(4),
s. 1364–1367.
doi:
10.4049/jimmunol.1500904.
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Huse, Kanutte; Bakkebø, Maren; Waelchli, Sebastien; Oksvold, Morten Pedersen; Hilden, Vera Irene & Forfang, Lise
[Vis alle 11 forfattere av denne artikkelen]
(2012).
Role of Smad Proteins in Resistance to BMP-Induced Growth Inhibition in B-Cell Lymphoma.
PLOS ONE.
7(10).
doi:
10.1371/journal.pone.0046117.
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Bone morphogenetic protein (BMP) expression and signaling are altered in a variety of cancers, but the functional impact of these alterations is uncertain. In this study we investigated the impact of expression of multiple BMPs and their signaling pathway components in human B-cell lymphoma. BMP messages, in particular BMP7, were detected in normal and malignant B cells. Addition of exogenous BMPs inhibited DNA synthesis in most lymphoma cell lines examined, but some cell lines were resistant. Tumor specimens from three out of five lymphoma patients were also resistant to BMPs, as determined by no activation of the BMP effectors Smad1/5/8. We have previously shown that BMP-7 potently induced apoptosis in normal B cells, which was in contrast to no or little inhibitory effect of this BMP in the lymphoma cells tested. BMP-resistance mechanisms were investigated by comparing sensitive and resistant cell lines. While BMP receptors are downregulated in many cancers, we documented similar receptor levels in resistant and sensitive lymphoma cells. We found a positive correlation between activation of Smad1/5/8 and inhibition of DNA synthesis. Gene expression analysis of two independent data sets showed that the levels of inhibitory Smads varied across different B-cell lymphoma. Furthermore, stable overexpression of Smad7 in two different BMP-sensitive cell lines with low endogenous levels of SMAD7, rendered them completely resistant to BMPs. This work highlights the role of Smads in determining the sensitivity to BMPs and shows that upregulation of Smad7 in cancer cells is sufficient to escape the negative effects of BMPs.
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Bakkebø, Maren Kolltveit; Huse, Kanutte; Hilden, Vera Irene; Smeland, Erlend B & Oksvold, Morten Pedersen
(2010).
TGF-beta-induced growth inhibition in B-cell lymphoma correlates with Smad1/5 signalling and constitutively active p38 MAPK.
BMC Immunology.
11.
doi:
10.1186/1471-2172-11-57.
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Sivertsen, Einar; Huse, Kanutte; Hystad, Marit Elise; Kersten, Christian; Smeland, Erlend B & Myklebust, June
(2007).
Inhibitory effects and target genes of bone morphogenetic protein 6 in Jurkat Tag cells.
European Journal of Immunology.
ISSN 0014-2980.
37(10),
s. 2937–2948.
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Huse, Kanutte
(2026).
Immune-cell-depleted DLBCLs have reduced expression of MHC class I.
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Lamont, Colin Harry; Nilsson, Daniel; Huse, Kanutte; Spasevska, Ivana; Olsson, Lina M. & Porwit, Anna
[Vis alle 10 forfattere av denne artikkelen]
(2026).
Tumor cell expression of Sox11 and cyclin D1 differently impact the spatial tumor immune cell landscape in Mantle cell lymphoma- a translational Nordic MCL2/3 study.
HemaSphere.
10(S1).
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Background
Mantle cell lymphoma (MCL) is a heterogeneous B cell lymphoma with frequent relapses. Intensified first line chemoimmunotherapy with cytarabine followed by autologous stem cell transplantation (ASCT) improves outcomes in younger patients, yet high risk disease (high Ki67/MIPI B and TP53 aberrations) remains difficult to control. The tumor microenvironment (TME) contributes to this heterogeneity as immune depleted, macrophage rich MCL have been linked to adverse prognosis and resistance. However, the spatial organization of TME and associated transcriptional programs in clinically annotated cohorts are incompletely defined.
Aims
To integrate spatial proteomics with compartment resolved transcriptomics to map TME architecture in MCL and relate it to tumor biology and outcome.
Methods
We included TMA sections containing 135 diagnostic tissue cores from 98 patients treated on Nordic Lymphoma Group trials MCL2/3 with long term follow-up clinical data. Imaging mass cytometry (IMC, Hyperion) used a 31 marker panel covering tumor, T cell, myeloid, endothelial and checkpoint proteins. GeoMx™ DSP profiled 1,811 transcripts (Cancer Transcriptomic Atlas) in 190 ROIs (~2 ROIs/patient), segmented into CD20+ tumor, CD8+ cytotoxic T cells, and CD3+/CD8− helper T cells.
Results
Fourteen distinct cell types/subsets were identifed in IMC data and unsupervised analyses of these cell type frequencies identified three recurrent TME clusters: tumor rich, M2 like macrophage rich, and T cell rich. Patients with M2 rich TME had inferior progression free survival and had tumor cells with lower SOX11 and higher Ki67 and cyclin D1 protein expression. Transcriptomics analysis of tumor cell segments revealed that SOX11 low tumors (enriched in the M2 rich cluster) had upregulated proliferation and cell cycle programs (e.g., Cyclin D1/Ki67 signatures), in contrast to SOX11+ tumor cells characterized by B cell differentiation/activation program and immune regulatory genes. Exhaustion associated markers (LAG3, TIGIT) were detected in T cells within M2 rich regions, yet accompanied by higher activation marker expression relative to the other clusters. Canonical correlation analysis highlighted the inverse relationship between cyclin D1 and SOX11, and SOX11 as the strongest tumor cell feature linked to neighborhood composition: SOX11 expression correlated positively with dendritic cells and PD 1+ CD8 T cells and negatively with PD 1− CD4 and CD8 T cells and macrophage abundance.
Conclusion
MCL exhibits distinct spatial TIME architectures with prognostic relevance. A macrophage dominant, SOX11 low state couples increased tumor proliferation with altered T cell phenotypes and inferior outcome. SOX11 emerges as a central axis, connecting tumor intrinsic programs to immune neighborhood structure, supporting its use for risk stratification and for guiding microenvironment targeted strategies (e.g., macrophage reprogramming and checkpoint/exhaustion pathway combinations) in high risk MCL.
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Kongsrud, Karoline; Tvedt, Charlotte Hop; Christensen, Thea Josefine; Caulier, Benjamin William; Spasevska, Ivana & Hilden, Vera Irene
[Vis alle 14 forfattere av denne artikkelen]
(2025).
B-cell Lymphoma Patient-Derived Xenograft Models for improved preclinical in vivo testing.
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The treatment landscape for B-cell non-Hodgkin lymphoma (B-NHL) has expanded dramatically in recent years, including advanced immunotherapies and targeted drugs. Yet, early relapse is still associated with an adverse prognosis, highlighting the need for new treatment options. For identification of potent new treatment strategies and combinatorial approaches, better preclinical models are needed that can facilitate testing at larger scale. Patient-derived xenograft (PDX) models are in vivo models established by engrafting tumor fragments from cancer patients into immunodeficient mice. The PDX tumors can retain the molecular features including clonal heterogeneity of the original tumor. Our aim is to establish relevant PDX models from B-cell lymphoma tumors for in vivo and ex vivo testing of drugs and immunotherapy.
We have successfully established PDX models from diffuse large B-cell lymphoma (DLBCL, n= 3) and mantle cell lymphoma (MCL, n= 2). For phenotypical characterization, we have developed a 32-marker spectral flow cytometry panel to identify key B cell features (CD10, CD19, CD20, CD22, CD37, CD79B, and Ig heavy and Ig light chains) and other immune cells including T cell subsets and myeloid cells. In addition, targeted sequencing will be used to determine if the mutational composition is stable over time, across different PDX generations. These results will facilitate selection of the best models for future studies. As proof-of -concept, we have used one of the MCL PDX models to test the efficacy of kinase inhibitors and the efficacy of CD19 CAR T cells. These models might support the development of personalized treatment strategies.
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Spasevska, Ivana; Kongsrud, Karoline; Sharma, Ankush; Scherer, Eveline; Hoel, Hanna Julie & Vincenti, Marianna
[Vis alle 11 forfattere av denne artikkelen]
(2025).
Beyond Helpers: Unraveling the Potential of CD4+ Cytotoxic T Cells in B cell lymphoma.
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CD4+ T cells are heterogeneous and consist of several subsets. While most CD4+ T cells exhibit either helper or immunoregulatory function, emerging evidence suggest the existence of a unique subset of CD4+ T cells with direct cytotoxic potential. Recently, there has been growing interest in these CD4+ cytotoxic T lymphocytes (CTLs) due to their potential involvement in anti-tumor immune responses and their relevance in immunotherapy.
We reanalyzed our prior single-cell RNA sequencing (scRNA-seq) dataset (Spasevska et al, Blood Adv 2023) to unravel the landscape of CD4+ T cell states in diffuse large B-cell lymphoma (DLBCL, n = 3), follicular lymphoma (FL, n = 3) and non-malignant tonsils (n = 3). Among 13 clusters, one was annotated CD4 cytotoxic (CTL) due to high expression of a cytotoxic gene signature (GZMK, NKG7, CST7, GZMA, GZMB). CD4+ CTLs represented over 20 % of CD4+ T cells in DLBCL and over 10% in FL tumors but were nearly absent in non-malignant tonsils. Their tumor-specific nature was further highlighted by a shared TCR observed in more than 45% of CD4+ CTLs in DLBCL. This suggests a clonal expansion of CD4+ T cells recognizing tumor antigens. To place the CD4 CTL in the context of other cytotoxic cells (CD8 and NK cells), we performed scRNA-seq of live cells from matched tumor and PBMC from two FL patients and one healthy donor PBMC (>50 000 single cells). Clustering and analysis of cytotoxic genes confirmed high similarity between CD4 CTL, CD8 Temra, CD8Tem and NK cells.
We validated these findings using multiparameter flow cytometry, revealing that CD4+ CTLs comprised from 1-23% of intratumoral CD4+ T cells in DLBCL. The CD4+ CTLs were identified as Granzyme B+ and Perforin+ cells within the CD4+ CD8- FOXP3- CD56- population, thereby excluding CD8+ T cells, regulatory T cells and NK cells. Using a 41-marker antibody panel for spectral flow cytometry, we have further validated that CD4+ CTL share a cytotoxic signature with NK cells and CD8 T cells. We will alto test if CD4 CTLs have comparable killing capacity to CD8 T cells by employing cell sorting and functional assays.
By employing state-of-the-art technologies, we have characterized CD4+ CTLs in lymphoma and demonstrated substantial clonal expansion in DLBCL. Our findings contribute to the understanding of immune cell heterogeneity and might have implications for therapeutic strategies that leverage the potential of CD4+ CTLs in cancer immunotherapy.
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Lamont, Colin Harry; Nilsson, Daniel; Huse, Kanutte; Spasevska, Ivana; Olsson, Lina M. & Kolstad, Arne
[Vis alle 10 forfattere av denne artikkelen]
(2025).
Tumor immune cell landscape of Mantle cell lymphoma.
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Mantle cell lymphoma (MCL) is a rare form of B-cell non-Hodgkin lymphoma (B-NHL). The malignancy commonly has an aggressive clinical course but is biologically and clinically heterogeneous. MCL overexpress cyclin D1 due to the t(11;14) translocation and most cases have aberrant expression of the neural transcription factor Sox11. Whereas the genetic landscape of MCL is well characterized, an in-dept characterization of the tumor immune microenvironment (TIME) is lacking. Here, we employed spatial proteomics to map the TIME architecture and explored how gene expression in tumor cells and T cells changed dependent on the neighborhood type. The cohort was based on TMA sections containing 134 diagnostic tumor tissues from 98 MCL patients included in the Nordic clinical phase II studies MCL2/MCL3. TMA neighbor sections were analyzed by imaging mass cytometry (IMC, Hyperion) and GeoMx (Nanostring DSP) was used to collect region of interest (ROI) spatially resolved genes expression profiles (immunopanel) from CD20+ cells (tumor), CD3+CD8+ T cells and CD3+CD8- T cells. By IMC, we identified 14 different cell types and 3 main TIME subtypes: T cell rich (47%), tumor rich (41%) and M2 rich (12%). The M2 rich subtype was enriched for cases with high protein expression of cyclin D1, low or negative expression of Sox 11 and higher gene expression of genes involved in DNA replication (MCM5, PLK1), MHC class I antigen presentation (TAP1) and interferon signaling (STAT1) in tumor cells. Sox11 expression was positively correlated with the cell ratios of DC/macrophages, PD-1+/PD1- CD8 T cells and PD-1+/PD-1- CD4 T cells, suggesting more antigen presentation to T cells in Sox11high tumors. Neighborhood analysis organizing MCL TIME into tumor, margin, follicle and T cell zones revealed that more PD-1+ CD4 T cells than PD-1+ CD8 T-cells co-localized to tumor zones. Survival analysis by applying univariate or multivariate proportional hazard analysis confirmed that patients with high M2/M1 macrophages had poor outcome whereas cases with remnant follicular zones had better outcome. Unsupervised integrative analysis using multi-omics factor analysis (MOFA), which included clinical, IMC and transcriptome data identified major dimensions of disease heterogeneity in MCL. Collectively, our data identified TIME subtypes and spatial features that refine disease heterogeneity and aggressiveness of MCL.
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Kared, Hassen; Huse, Kanutte; Spasevska, Ivana; Myklebust, June Helen & Munthe, Ludvig Andre
(2025).
Mapping T Cell subsets and TME types in Prostate Cancer.
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Sharma, Ankush; Kongsrud, Karoline; Spasevska, Ivana; Lamont, Colin Harry; Steen, Chloe Beate & Lorenz, Susanne
[Vis alle 10 forfattere av denne artikkelen]
(2025).
Deciphering Tumor Microenvironment Complexity in Lymphoma Using Spatial Transcriptomics.
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The tumor microenvironment (TME) in lymphomas, such as Diffuse Large B-cell Lymphoma (DLBCL) and Follicular Lymphoma (FL), is highly heterogeneous and plays a critical role in modulating therapeutic outcomes, including responses to CD19 CAR T-cell therapy. To unravel the spatial complexity of the TME, we employed an integrative approach combining spatial transcriptomics (Visium, 10x Genomics) and spatial proteomics (imaging mass cytometry, IMC) on FFPE tissue sections from FL and DLBCL patients. Using tools such as CytoSPACE and CARD, we mapped single-cell RNA-seq data to spatial coordinates and validated cellular compositions with IMC data. We identified spatially distinct immune and stromal cell populations, including GC B cells, Tregs, macrophages, dendritic cells, and endothelial cells, across follicular and T-cell rich regions. CytoSPACE-based mapping revealed consistent enrichment of key markers (e.g., CD79B, FOXP3, GRZA) with corresponding protein expression profiles. We also developed R based ShinySpatial framework to visualize spatial data enhancing the accessibility and interpretability of complex spatial datasets. This pilot study lays the groundwork for identifying spatial biomarkers that predict therapeutic responses and resistance in lymphoma, with ongoing integration of data from patients enrolled in the JULIET, BELINDA, and ELARA trials.
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Lamont, Colin Harry; Huse, Kanutte; Spasevska, Ivana; Nilsson, Daniel; Kolstad, Arne & Jerkeman, Mats
[Vis alle 8 forfattere av denne artikkelen]
(2024).
Mapping the tumor microenvironment in mantle cell lymphoma by imaging mass cytometry.
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Background: Mantle cell lymphoma (MCL) is a rare aggressive form of B cell non-Hodgkin lymphoma, accounting for less than 7% of all cases of newly diagnosed lymphoma. MCL is characterized by specific hallmarks, including the t(11;14)(q13;q32) translocation that results in constitutive expression of Cyclin D1, and overexpression of the SOX11 transcription factor. Although the molecular profile and the genetic landscape of MCL and their impact on the disease aggressiveness have been well characterized, the tumor microenvironment (TME) and its role are still not fully understood. Here, we aimed to explore TME in MCL, determine how tumor cell features shape the TME and link TME features to clinical high-risk factors.
Methods: Diagnostic biopsies from 102 MCL patients included in the two 2 prospective trials by the Nordic Lymphoma Group, MCL2 and MCL3, were imaged by Hyperion imaging mass cytometry (IMC) using a 31-marker antibody panel. Images were segmented into single cells, batch-corrected and annotated using a novel binary clustering strategy. Using CytoMAP, the cells were divided into 30 µm-radius neighborhoods and clustered to define five types of tissue regions.
Results: We identified 13 different clusters of cells present in the MCL TME, with CD4 T cells being the most prevalent immune cell population, with a median of 10% (range 2.2-28%), followed by CD8 T cells with a median of 9% (range 1.5-22%) and CD163- macrophages 8% (range 2.1-23%). T follicular helper cells are notably absent and the follicular dendritic cell (FDC) networks are heterogenous. Neighborhood analysis reveals five types of microenvironments across all patients: tumor exclusive (22%), immune-infiltrating tumor (31%) immune-dominant tumor (19%), T cell zone like (6%), and FDC/follicle (14%). Hierarchical clustering of patients reveals that differences in the microenvironment composition are associated with architectural motifs and follicle organization.
In our dataset, 89 patients were SOX11+, and high expression of SOX11 was associated with higher CD19 (p = 0.03, two sided Mann-Whitney test) and lower Cyclin D1 (p = 2x10⁻⁴) and Ki67 (p = 1.7 x10⁻³) expression and linked to a longer time to progression (p = 0.04).
Conclusion: Our study contributes to an improved understanding of the MCL tumor microenvironment. Future analysis will determine how the observed tumor microenvironment shapes clinical outcomes.
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Huse, Kanutte; Bakkebø, Maren; Forfang, Lise; Hilden, Vera Irene; Smeland, Erlend B & Myklebust, June
(2010).
Bone Morphogenetic Proteins Inhibit CD40L-and IL-21-Induced Ig Production In Human B Cells: Differential Effects of BMP-6 and BMP-7.
Blood.
ISSN 0006-4971.
116(21),
s. 729–729.
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Publisert
4. apr. 2023 13:17
- Sist endret
5. mai 2023 13:08
