Undefined Profile Research Articles

Abstract 3708: Characterization of EpCAM-positive cells for specific detection of disseminated cancer cells in bone marrow of early breast cancer patients

Abstract Obviously, breast cancer cells that disseminated from the primary tumor before surgical resection are responsible for metastasis. However, detection of disseminated cancer cells (DCC) in breast cancer patients during minimal residual disease is still difficult, impeding the study of early systemic cancer and the extraction of clinically relevant diagnostic or therapeutic information. Others and we have shown previously that EpCAM is a suitable DCC detection marker, however not entirely specific in bone marrow. To improve DCC detection, we here compared different methods to enrich DCC exploiting information from our ongoing single cell RNA-seq study. Based on our ongoing scRNA-seq study (see poster of Elia Raya), we generated a qPCR assay to classify candidate DCC based on an expression signature of four genes. This assay categorizes plasma cell-like EpCAM+ cells from healthy donors and candidate DCC from breast cancer patients with high fidelity. With this assay at hand, we compared two enrichment strategies: depletion of CD11b, CD319 and Glycophorin A positive cells from density gradient enriched mononuclear cells (MNC) vs. positive selection of EpCAM+ cells and subsequent addition of markers that separate out DCC among EpCAM+ cells. For this we applied 10x genomics profiling of EpCAM+ cells from bone marrow. We depleted and screened the bone marrow of 221 patients suffering from non-metastatic early breast cancer for EpCAM-positive cells. From 87 patient samples we could isolate EpCAM-positive cells. We performed single cell RNA-sequencing on 64 EpCAM-positive cells from 54 patients of which 84% were classified as cancer cells and 16% of bone marrow origin. Applying the selected gene signature, 76% of isolated cells were classified as DCC, 16% were undefined and 8% as non-cancer bone marrow cell (NCC). Upon positive selection of EpCAM+ cells implementing additional markers, we isolated 53 cells from 14 patients. Of these, 68% showed a DCC or DCC-like-signature, 15 % a NCC and 17% an undefined profile. To get a better understanding of these EpCAM-positive confounder populations we currently perform a scRNA-seq profiling using 10X on the enriched EpCAM+ cells. Our data shows that EpCAM is a promising marker to identify DCCs in the bone marrow of early cancer patients, but it also indicates that we find several groups of confounder cells. To differentiate between these cell types during staining and in our qPCR assay we are currently testing additional markers. Citation Format: Jonas Roth, Elia Raya, Sarah Hücker, Huiqin Koerkel-Qu, Stephan Seitz, Christoph Klein. Characterization of EpCAM-positive cells for specific detection of disseminated cancer cells in bone marrow of early breast cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3708.

Human macrophages chronically exposed to LPS can be reactivated by stimulation with MDP to acquire an antimicrobial phenotype

Macrophages are important in host defense and can differentiate into functionally distinct subsets named classically (M1) or alternatively (M2) activated. In several inflammatory disorders, macrophages become tolerized to prevent deleterious consequences. This tolerization reduces the ability of macrophages to respond to bacterial components (e.g., LPS) maintaining a low level of inflammation but compromising the ability of macrophages to mount an effective immune response during subsequent pathogen encounters.In this study, we aimed to reactivate human monocyte-derived macrophages chronically exposed to LPS by re-stimulation with muramyl dipeptide (MDP). We observed an undefined profile of cell surface marker expression during endotoxin tolerance and absence of TNFα production. Stimulating macrophages chronically exposed to LPS with LPS+MDP restored TNFα, production together with an increased production of IL1, IL6, IFNγ, IL4, IL5 and IL10. These results suggest that macrophages chronically exposed to LPS possess a mixed M1-M2 phenotype with sufficient antimicrobial and homeostatic potential.

Exploring the Interplay of Adaptive and Maladaptive Strategies

In emotion regulation research, it is common to distinguish adaptive from maladaptive emotion regulation strategies. However, little is known about their interactional impact (compensational or interfering effects) on adolescents’ adjustment. We collected longitudinal, multiple informant questionnaire data from N = 608 adolescents and their parents to examine the prevalence of adolescents’ anger regulation profiles using latent profile analysis. We identified five anger regulation profiles—an adaptive, a maladaptive, a low, a medium, and an undefined profile. The first three profiles could be replicated 1 year later, as could one additional multiple and another undefined profile. Findings supported the assumption that adaptive strategies compensate for the negative impact of maladaptive strategies, yielding negative psychosocial consequences, particularly for the maladaptive profiles. Moreover, different profiles were divergently related to adolescents’ internalizing and externalizing problems concurrently and over time. Results highlight the importance of considering both adaptive and maladaptive anger regulation strategies in prevention and psychotherapy.

Isolated anti‐β2‐glycoprotein I antibodies in neurology: a frontier syndrome between multiple sclerosis and antiphospholipid syndrome?

Anti-β2-glycoprotein I (anti-β2-GPI) antibodies are part of the heterogeneous family of antiphospholipid antibodies and seem to be present in various neurological manifestations in addition to antiphospholipid syndrome (APS). Our objective was to analyse the clinical, radiological and therapeutic characteristics of neurological patients with positive anti-β2-GPI antibodies and without the Sapporo criteria for APS. The medical records were retrospectively reviewed of 28 consecutive patients hospitalized in the Neurology Department of Strasbourg University Hospital, France, in whom anti-β2-GPI antibodies (immunoglobulin G and/or immunoglobulin M) were positive and other antiphospholipid antibodies negative, from November 2005 to July 2011. Clinical, radiological, biological and therapeutic data and clinical course were studied. Positive anti-β2-GPI antibodies were present in 28 patients. The predominant physiopathological process was mainly inflammatory (25% with myelitis, 14.3% with optic neuritis) or vascular (14.3% with cerebral ischaemia, 7.1% with cerebral vasculitis). Brain magnetic resonance imaging was performed in 89.3% of patients: atypical lesions were observed in 44% and typical inflammatory and vascular lesions in 16% and 12%, respectively. The anti-β2-GPI antibody seems to be involved in two types of neurological disease: vascular or inflammatory 'multiple sclerosis-like' disease. These two types of patients frequently develop an autoimmune disease (multiple sclerosis, systemic lupus erythematosus, APS). However, a large proportion of the patients had an undefined profile with aspecific cerebral lesions and required monitoring. This study raises questions about a separate entity at the border between APS and multiple sclerosis which remains to be better defined in a larger cohort.

Defining undifferentiated connective tissue diseases: a challenge for rheumatologists

Undifferentiated connective tissue diseases: distinct clinical entities? Classification criteria have been developed by the scientific community to improve communication and facilitate clinical studies. In clinical practice, however, it is not unusual to come across systemic autoimmune diseases that do not meet the existing classification criteria. These diseases have been variously termed as incomplete lupus erythematosus, undifferentiated connective tissue diseases (UCTD) or undifferentiated connective tissue syndromes. 1–18 However, a review of the literature shows that a broad range of conditions including transitory diseases, the initial phases of defined CTD, incomplete forms of defined CTD, and conditions that cannot be classified or diagnosed based on the clinical findings have been classified as UCTD. 4,9,15,19–24 Studies demonstrate that a certain proportion of UCTD patients will eventually develop defined CTD, generally systemic lupus erythematosus (SLE), but others develop conditions such as systemic sclerosis (SSc), Sjogren’s syndrome, mixed connective tissue diseases, rheumatoid arthritis (RA), systemic vasculitis and polydermatomyositis. 2–4,6–9,11–15 The marked discrepancy in the reported rates of such an evolution – ranging from 10% to 70% – may be attributable to the characteristics of the patients recruited in different studies, in particular their disease duration. Higher incidences have been reported during the first 3– 5 years of the disease, and, therefore, studies that include patients with recent onset UCTD are likely to see a larger number evolving to a CTD. In contrast, UCTD of long duration is much less likely to develop into a defined CTD. At the same time, a distinct subgroup of patients can be identified who will maintain an undefined profile during follow-up. 4,6–8,11–15 Their disease can be termed stable UCTD, a condition with low disease activity, a low incidence of flares (7% in our patient population over 1 year; unpublished data) and the absence of severe organ involvement (e.g. pulmonary fibrosis, kidney or neurological impairment). The most common clinical manifestations are non-erosive arthritis; Raynaud’s phenomenon without swollen hands, sclerodactily or nailfold capillaroscopic abnormalities suggestive of SSc; sicca symptoms (but negative salivary gland biopsy and ocular test results) and leucopenia. The serological profile is positive for antinuclear antibodies (ANA), generally with a single autoantibody specificity (anti-Ro/SSA or anti-RNP) and only rarely disease-specific autoantibodies (usually anti

High dispersion spectroscopic study of quiescent prominences

The utility of very high dispersion spectra (5–11 mm/A) for the study of line profile and velocity structure in quiescent prominences is demonstrated by observations, taken with the spectregraphic slit positioned normal to the limb in Hα λ6563 A, He D3 λ5876 A, and Ca+K λ3933 A. The emission profiles of both Hα and the K line often show a central reversal (absorption). Emission structures in the K-line can be complex with details as narrow as 0.04 A. Frequently this structure consists of two distinct components: a central, strong, rather narrow line, and an often displaced, weak feature of undefined profile appearing as ‘fuzz’. It is suggested that this fuzz indicates an exchange of matter between the prominence and the corona.