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Article

HMGB1 Deficiency Occurs in a Broad Range of Human Cancers and Is Often Associated with Unfavorable Tumor Phenotype

by
Viktoria Chirico
1,
Hena Sharifi
1,
Maria Christina Tsourlakis
1,
Seyma Büyücek
1,
Clara Marie von Bargen
1,
Katharina Möller
1,
Florian Lutz
1,
David Dum
1,
Martina Kluth
1,
Claudia Hube-Magg
1,
Georgia Makrypidi-Fraune
1,
Piero Caneve
1,
Maximilian Lennartz
1,
Morton Freytag
1,
Sebastian Dwertmann Rico
1,
Simon Kind
1,
Viktor Reiswich
1,
Eike Burandt
1,
Till S. Clauditz
1,
Patrick Lebok
1,2,
Christoph Fraune
1,2,
Till Krech
1,2,
Sarah Minner
1,
Andreas H. Marx
3,
Waldemar Wilczak
1,
Ronald Simon
1,*,
Guido Sauter
1,
Stefan Steurer
1 and
Kristina Jansen
4add Show full author list remove Hide full author list
1
Institute of Pathology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
2
Institute of Pathology, Clinical Center Osnabrück, 49074 Osnabrück, Germany
3
Department of Pathology, Academic Hospital Fürth, 90766 Fürth, Germany
4
General, Visceral and Thoracic Surgery Department and Clinic, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
*
Author to whom correspondence should be addressed.
Diagnostics 2025, 15(15), 1974; https://doi.org/10.3390/diagnostics15151974
Submission received: 11 April 2025 / Revised: 30 July 2025 / Accepted: 30 July 2025 / Published: 6 August 2025
(This article belongs to the Section Pathology and Molecular Diagnostics)
Browse Figures
Versions Notes

Abstract

Background/Objectives: Aberrant expression of high-mobility group protein B1 (HMGB1) has been linked to cancer development and progression. Methods: To better comprehend the role of HMGB1 expression in cancer, a tissue microarray containing 14,966 samples from 134 different tumor entities and 608 samples of 76 different normal tissue types was analyzed by immunohistochemistry. Results: Strong HMGB1 staining occurred in almost all normal cell types and in most cancers. Of 11,808 evaluable cancers, only 7.8% showed complete absence of HMGB1 staining (HMGB1 deficiency) while 9.9% showed 1+, 25.0% showed 2+, and 57.2% showed 3+ HMGB1 positivity. Absence of HMGB1 staining mostly occurred in pheochromocytoma (90.0%), seminoma (72.4%), gastrointestinal stromal tumor (28.6%), adrenal cortical carcinoma (25.0%), and Hodgkin’s lymphoma (25.0%). Low HMGB1 staining was linked to poor histologic grade (p < 0.0001), advanced pT stage (p < 0.0001), high UICC stage (p < 0.0001), and distant metastasis (p = 0.0413) in clear cell renal cell carcinoma, invasive tumor growth in urothelial carcinoma (pTa vs. pT2–4, p < 0.0001), mismatch repair deficiency (p = 0.0167) in colorectal cancers, and advanced pT stage in invasive breast carcinoma of no special type (p = 0.0038). Strong HMGB1 staining was linked to nodal metastases in high-grade serous ovarian carcinomas (p = 0.0213) and colorectal adenocarcinomas (p = 0.0137), as well as to poor histological grade in squamous cell carcinomas (p = 0.0010). Conclusions: HMGB1 deficiency and reduced HMGB1 expression occur in a broad range of different tumor entities. Low rather than strong HMGB1 staining is often linked to an aggressive tumor phenotype. Whether HMGB1 deficiency renders cells susceptible to specific drugs remains to be determined.

1. Introduction

High-mobility group protein B1 (HMGB1) is the second most common protein after histone in the nucleus of most human cell types. Like all other members of the non-histone chromosomal high-mobility group protein family, HMGB1 is a chromatin-associated protein. It has a role in the maintenance of nucleosome structure, regulation of DNA replication, transcription control, and DNA repair through a specific binding to damaged DNA. HMGB1 increases the binding affinity of many transcription factors, such as p53, Rb, NF-κB, and estrogen receptor, to their target DNA sequences [1]. Critical additional functions of HMGB1 require an extra-nuclear location of the protein. In the cytoplasm, HMGB1 participates in the control of autophagy and apoptosis [2,3]. After release to the extracellular space, HMGB1 acts as a damage-associated molecular pattern molecule (DAMP) that critically mediates inflammation and immune responses in various conditions, including sepsis, atherosclerosis, arthritis, neurodegeneration, meningitis, and cancer. Therapeutic options to regulate HMGB1 in preclinical models are being evaluated in the field of non-neoplastic diseases (sepsis, inflammation) [4], as well as in cancer [5,6,7,8,9].
The role of HMGB1 in cancer is complex and not well understood. Different mechanisms activated by HMGB1 contribute to critical cancer cell traits such as autophagy, immunogenic cell death (ICD), release of cytokines and chemokines, angiogenesis, and migration [10,11,12]. Both increased and reduced levels of HMGB1 in cancer cells have been found to be linked to aggressive cancer phenotype, and it was proposed that these different roles may depend on the tissue/cell types involved [13].
In case of overexpression, several pivotal transcription factors, such as p53, retinoblastoma (RB) proteins, and NF-κB family members, can enhance their oncogenic activities through direct interactions with HMGB1 [14,15,16]. Reduced levels of expression or activity of HMGB1 in cancer cells may result in an unfavorable disease course due to increased levels of genomic instability [17,18]. The fact that HMGB1 is expressed not only in cancer cells but also in virtually all cell types of the tumor microenvironment adds further complexity and constitutes a challenge for studying HMGB1 in cancer in vivo. Although immunohistochemistry (IHC) enables a cell type and cell compartment-specific evaluation of HMGB1 protein, the number of previous studies is limited, and the obtained data are difficult to compile because of different antibodies, staining protocols, and evaluation criteria that were used in these studies.
We hypothesized that HMGB1 alterations may correlate with more aggressive clinicopathological features in different tumor types. Accordingly, more than 14,000 tissue samples from 134 different tumor types and subtypes, as well as 76 non-neoplastic tissues, were evaluated by IHC in a tissue microarray (TMA) format. Results were compared with histological parameters of malignancy, clinical outcome in some tumor entities, as well as with previously collected data on the immune cell tumor microenvironment.

2. Materials and Methods

2.1. Tissue Microarrays (TMAs)

Our normal tissue TMA was composed of 8 samples from 8 different donors for each of 76 different normal tissue types (608 samples on one slide). The cancer TMAs contained a total of 14,966 primary tumors from 134 tumor types and subtypes. Detailed histopathological and molecular data were available for cancers of the kidney (n = 1757), urinary bladder (n = 829), colorectum (n = 2351), pancreas (n = 598), stomach (n = 327), liver (n = 301), ovary (n = 524), and the thyroid gland (n = 518). Data on pT, pN, grade, and HPV status were available from 902 squamous cell carcinomas of 11 different sites of origin. The composition of both normal and cancer TMAs is described in detail in Section 3. All samples were obtained from the archives of the pathology institutes in Hamburg, Osnabrück, and Fürth (University Medical Center Hamburg-Eppendorf, Clinical Center Osnabrück, Academic Hospital Fürth, Germany). Tissues were fixed in 4% buffered formalin and then embedded in paraffin. TMA tissue spot diameter was 0.6 mm. The use of archived remnants of diagnostic tissues for the manufacturing of TMAs and their analysis for research purposes, as well as patient data analysis, is permitted in accordance with local laws (HmbKHG, §12,1) and has been approved by the local ethics committee (Ethics commission Hamburg, WF-049/09). All work has been carried out in compliance with the Helsinki Declaration.

2.2. Immunohistochemistry

Freshly cut TMA sections were immunostained on one day and in one experiment. Slides were deparaffinized with xylol, rehydrated through a graded alcohol series, and exposed to heat-induced antigen retrieval for 5 min in an autoclave at 121 °C in pH 7.8 Tris-EDTA-Citrate (TEC) buffer. Endogenous peroxidase activity was blocked with Dako REAL Peroxidase-Blocking Solution (Agilent Technologies, Santa Clara, CA, USA; #S2023) for 10 min. Primary antibody specific for HMGB1 (recombinant rabbit monoclonal, clone HMV317, cat. #6559-317-01, MS Validated Antibodies GmbH, Hamburg, Germany) was applied at 37 °C for 60 min at a dilution of 1:150. This immunostaining protocol was developed using a small test TMA containing negative (spermatids in the testis) and positive (colon mucosa) control tissues. The normal tissue TMA (NTA) was also analyzed with an independent antibody against HMGB1 (EPR3507, Abcam, Cambridge, U.K., cat. #ab79823, dilution 1:3600) to validate the specificity of HMV317 by comparing the staining patterns of both antibodies as suggested by the International Working Group for Antibody Validation (IWGAV) [19]. According to the IWGA, specificity can be assumed when two independent antibodies show identical staining, while all staining observed only with one but not with the other antibody must be considered antibody-specific cross-reactivity. The NTA, with its broad range of human healthy tissues, is optimally suited to detect antibody-specific cross-reactivity as it represents virtually all human proteins. Examples for the comparison of the two antibodies are provided in Supplementary Figure S1. Bound antibody was then visualized using the Dako REAL EnVision Detection System Peroxidase/DAB+, Rabbit/Mouse kit (Agilent Technologies, Santa Clara, CA, USA; #K5007) according to the manufacturer’s directions. The sections were counterstained with hemalaun. One experienced pathologist (V.C.) analyzed all tumor TMA sections according to an established scoring system [20]. In brief, the staining intensity of tumor cells was semi-quantitatively recorded as 0, 1+, 2+, or 3+. Presence of unequivocal HMGB1 positivity in stroma cells was particularly required to classify a tumor as “0” (HMGB1 completely negative, HMGB1 deficiency). Tumors with the absence of HMGB1 staining in both tumor and stroma cells were categorized as “non-informative”. This scoring scheme was selected to allow for unequivocal identification of tumors with HMGB1 deficiency, for statistical comparison between tumors with and without HMGB1 deficiency, and between tumors with low (1+), high (3+), or intermediate (2+) HMGB1 expression Supplementary Figure S2.

2.3. Statistics

Statistical calculations were performed with JMP® version 18 software (SAS®, Cary, NC, USA). Contingency tables and the chi2-test were performed to search for associations between HMGB1 immunostaining and tumor phenotype.

3. Results

3.1. Technical Issues

A total of 11,808 (78.9%) of 14,966 tumor samples were interpretable in our TMA analysis. Non-interpretable samples demonstrated a lack of unequivocal tumor cells, a lack of the entire tissue spot, or a lack of staining in neoplastic and normal cells. A sufficient number of samples (≥4) of each normal tissue type was evaluable.

3.2. HMGB1 in Normal Tissues

A distinct, and usually strong, nuclear HMGB1 staining was observed in all tissues and in the vast majority of cell types. HMGB1 staining was particularly strong in lymphocytes and other cells of the immune system, all cells of the hematopoietic system, epithelial cells of the gastrointestinal tract, pneumocytes, epithelial cells of the fallopian tube, breast epithelial cells, epithelial and stromal cells of the proliferating endometrium, ovarian stroma cells, epithelial cells of the parathyroid gland, and in granule cells of the cerebrum. In squamous epithelium, HMGB1 staining was strong in the basal and suprabasal cell layers, but the staining intensity continuously decreased towards the superficial cell layers, where the staining sometimes was faint. Nuclear HMGB1 labeling was also rather weak in central areas of corpuscles of Hassall’s of the thymus, hepatocytes, Brunner glands, proximal tubuli of the kidney, acinar cells of the prostate (while it was markedly stronger in basal cells), and in corpus luteum cells of the ovary. Nuclear HMGB1 staining was—under the selected conditions—faint or absent in heart muscle, epithelial cells of the adenohypophysis, pituicytes of the neurohypophysis, and in maturing germ cells of the testis (spermatocytes, spermatids). HMGB1 staining was distinct in cortical cells but only weak or even absent in medullary cells. Representative images are shown in Figure 1. All these stainings, including the distinction between low- and high-expressing cells, were obtained with both the HMV317 and the EPR3507 antibody (Supplementary Figure S1).

3.3. HMGB1 in Tumor Tissues

A nuclear HMGB1 staining was seen in all cells of the vast majority of tumors. Of 11,808 evaluable tumors, only 921 (7.8%) showed a complete lack of HMGB1 expression (staining intensity “0”, HMGB1 deficiency) while 1172 (9.9%) showed 1+, 2956 (25.0%) showed 2+, and 6759 (57.2%) showed 3+ HMGB1 positivity (Table 1).
HMGB1 deficiency most commonly occurred in pheochromocytoma (90.0%), seminoma (72.4%), gastrointestinal stromal tumor (28.6%), adrenal cortical carcinoma (25.0%), Hodgkin’s lymphoma (25.0%), Leydig cell tumor of the testis (16.7%), prostatic adenocarcinoma (8.9–16.4%), renal cell tumors (8.4–16.2%), yolk sac tumor of the testis (15.4%), paraganglioma (12.5%), neuroendocrine tumor of the lung (11.1%), adrenocortical adenoma (10.8%), epithelioid mesothelioma (10.7%), and in clear cell carcinoma of the ovary (9.8%). HMGB1 deficiency in <10% was seen in 52 other tumor categories. Representative images are shown in Figure 2, and other tumor entities are in Supplementary Figure S3.
A ranking order of tumors according to their rate of HMGB1 deficiency is given in Figure 3.
The comparison with clinico-pathological parameters of a malignant tumor phenotype revealed significant associations between altered HMGB1 expression and aggressive tumor phenotype in multiple tumor entities (Table 2).
Weak HMGB1 immunostaining was linked to poor histological grade (p < 0.0001), advanced pT stage (p < 0.0001), high UICC stage (p < 0.0001), nodal metastases (p = 0.0108) and distant metastasis (p = 0.0413) in clear cell renal cell carcinomas (ccRCC), invasive tumor growth in urothelial carcinoma (pTa vs. pT2–4, p < 0.0001), mismatch repair deficiency (p = 0.0167) in colorectal cancers, and advanced pT stage in invasive breast carcinomas of no special type (NST; p = 0.0038). Strong HMGB1 immunostaining was linked to nodal metastases in high-grade serous ovarian carcinomas (p = 0.0213) and colorectal cancers (p = 0.0137), as well as to high histological grade in a combined analysis of squamous cell carcinomas originating from 11 different organs (p = 0.0010). HMGB1 immunostaining was unrelated to parameters of cancer aggressiveness in pancreatic and gastric adenocarcinomas, seminomas, as well as in endometrioid endometrial carcinomas.

4. Discussion

The successful IHC analysis of 11,808 tumors from 134 different entities, together with a broad range of different normal tissues, provides a comprehensive overview of the pattern of HMGB1 expression in human tumors. The approximately 3000 non-analyzable TMA spots correspond to the typical rate of approximately 10–20% non-interpretable tissue spots in TMA studies [21,22]. Since the causes (missing spots or absence of tumor cells) can affect any spot of any tumor type at random, a general bias for the study results can be excluded. That a complete loss of HMGB1 expression (HMGB1 deficiency) can be found in at least one individual tumor from more than 75 different tumor categories represents a major finding of this study. The ranking list of tumors according to their rate of HMGB1 deficiency (Figure 3) provides probably the most comprehensive view of this alteration available to date, although the accuracy of our data is still limited by small numbers in some tumor categories. It is of note that seminoma and pheochromocytoma, the tumors with—by far—the highest rate of cases with HMGB1 deficiency, are derived from HMGB1-negative cell types. In these tumors lack of HMGB1 expression may represent a ”normal” status, while a lack of HMGB1 protein will indicate “HMGB1 deficiency” in most or all other tumor entities. In line with our data, others described HMGB1 deficiency in tumor cells to occur in subsets of colorectal [23] and ovarian cancers [24]. The large size of several of our tumor cohorts enabled us to address the clinical relevance of HMGB1 deficiency in various cancer types. The significant associations between HMGB1 deficiency and dismal tumor phenotype in ccRCC, urothelial carcinoma, and invasive breast carcinomas of no special type (NST) argue for low HMGB1 expression levels, going along with aggressive cancer behavior at least in certain cancer entities. Significant associations between HMGB1 deficiency in tumor cells and unfavorable tumor features have also been previously reported in pancreatic cancer [25] and endometrial cancer [26].
The molecular mechanisms resulting in reduced or lost HMGB1 expression and increased aggressiveness of HMGB1-depleted cancer cells are not fully understood. It has been shown, however, that genetic deletions [27], promoter hypermethylation [28], and microRNA-mediated suppression [29] can contribute to HMGB1 loss, and that HMGB1 deficiency can affect both tumor cell biology and tumor microenvironment. In line with an important role of HMGB1 for maintenance of genomic integrity, mouse fibroblasts lacking HMGB1 suffer from pronounced chromosomal instability and display higher rates of damage after UV irradiation than wildtype controls [17]. In pancreatic models, HMGB1 loss leads to oxidative DNA damage, chromosomal rearrangements, and telomere abnormalities, resulting in inflammatory nucleosome release and the propagation of KRAS-driven tumorigenesis [25]. Moreover, it has been shown that HMGB1-deficient tumors resist DNA-alkylating therapies and have an impaired ability to recruit immune cells into the treated tumor tissue, resulting in impaired apoptosis and improved cancer cell survival [30]. Although it cannot be excluded that loss of HMGB1 in cells of advanced tumors does not represent a functionally significant modification but just reflects tumor cell dedifferentiation, which typically parallels cancer progression, it appears counterintuitive that loss of the most abundant histone protein in cancer cells remains without a notable functional effect.
The complete loss of a protein with an important cellular function in tumor cells may offer therapeutic options targeting proteins from associated signaling pathways, such as DNA damage or innate immune response. In the case of HMGB1 deficiency, potential strategies have been proposed, although clinical trials are lacking to date. Since loss of HMGB1 in cancer cells leads to impaired DNA repair in vitro and in vivo, it has been suggested that HMGB1 deficiency may increase sensitivity to alkylating agents or ionizing radiation [31,32]. Moreover, the combination of chemotherapy and Toll-like receptor 4 (TLR4) agonists has been proposed as a potentially promising approach. Because HMGB1-deficient cancers may have limited capacity to recruit inflammatory cells through secretion of HMGB1, it was speculated that TLR4 agonists may be able to substitute HMGB1 in its role of activating T cells by binding to TLR4. One earlier study had shown that the use of TRL4 agonists promoted immunogenic cell death and may have helped to overcome the resistance observed in HMGB1-deficient tumors [33]. Irrespective of a possible type of action, our ranking order of tumors according to their rate of HMGB1 deficiency describes the tumor entities that might benefit most from potential future drugs for targeting HMGB1-deficient tumors.
Several previous IHC studies have reported associations between high HMGB1 immunostaining and unfavorable tumor phenotype or poor prognosis in non-small-cell lung cancer [34], cervical cancer [35], squamous cell carcinoma of the esophagus [36], squamous cell carcinoma of the larynx [37], urothelial carcinoma of the bladder [38], breast cancer [39], as well as colorectal [27] and gastric adenocarcinoma [40,41,42]. Our data revealed only comparatively weak associations between increased HMGB1 expression and unfavorable tumor features in serous ovarian carcinoma, colorectal cancers, and squamous cell carcinomas. It is of note, however, that our IHC protocol was designed to be highly sensitive by increasing the antibody concentration to the highest possible level before disturbing background staining occurred. Such a protocol ensures that even very low expression is reliably detected but also results in a loss of discrimination at high expression levels. As a result, the group of 3+ cases was quite large in most of our tumor categories. It cannot be excluded that a markedly less sensitive protocol would have distinguished a subgroup of tumors with a particularly high HMGB1 expression within our group 3+ positive tumors, and that these tumors could be characterized by increased aggressiveness. Moreover, due to low numbers of cases, associations with tumor phenotype could not be evaluated in this study for several cancer types (non-small-cell lung cancer, cervical cancer, gastric adenocarcinoma) for which an oncogenic role of HMGB1 has previously been described [27,36,37,38,39,40,41,42]. An oncogenic role of HMGB1—at least in specific tumor types—can therefore not be excluded.
Because of the ubiquitous expression of HMGB1 in virtually all cell types of the tumor stroma, IHC is the optimal method for assessing the HMGB1 status of a tumor. Automated image analysis tools could even enhance IHC scoring in specific tumor types, but algorithms that would enable the distinction between tumor and non-neoplastic cells across a broad range of different tumor types are currently lacking.
Considering the high variability of published IHC data on HMGB1, a particular emphasis was placed on the validation of our IHC assay. The International Working Group for Antibody Validation (IWGAV) has proposed that an acceptable antibody validation for IHC on formalin-fixed tissues must include either a comparison of IHC findings with a second antibody for the same target or a comparison with another independent method for expression analysis [19]. Because a comparison with data obtained by a method using disaggregated tissue is inappropriate in the case of ubiquitously expressed proteins, a thorough comparison with an independent second antibody was performed on consecutive TMA sections containing samples from 76 different normal tissue categories. The specificity of our assay was confirmed because identical staining patterns were observed with both HMV317 and EPR3507 antibodies, particularly in these tissues, where the staining displayed a variability between different cell types. These included a decrease in the staining intensity from basal to superficial cell layers in squamous epithelium and the absence of HMGB1 in maturing spermatids and spermatozoa of the testis and in medullary cells of the adrenal gland. A very broad range of different tissues for antibody validation increases the likelihood of detecting antibody cross-reactivities because virtually all proteins occurring in normal cells of adult humans are subjected to the validation experiment.
In summary, our data show that HMGB1 deficiency and reduced HMGB1 expression occur in a broad range of different tumor entities and that low rather than high HMGB1 expression is often linked to an aggressive tumor phenotype, thereby bridging the gap between basic molecular insights and clinical applications in cancer. If drugs became available that target HMGB1 deficiency, immunohistochemical testing would be suitable to identify patients who could potentially benefit from such therapies. However, it remains to be clarified whether HMGB1 deficiency makes the cells susceptible to certain drugs, whether HMGB1 deficiency can be used as a prognostic marker, and which thres-hold values are clinically applicable to define an immunohistochemical HMGB1 deficiency. In view of the retrospective nature and the limited number of cases of some tumor types, further studies for prospective validation in patient cohorts with follow-up data would be desirable.

Supplementary Materials

The following supporting information can be downloaded at https://www.mdpi.com/article/10.3390/diagnostics15151974/s1, Supplementary Figure S1. Immunohistochemistry validation by comparison with an independent second antibody. The panels show a full confirmation of IHC results by an independent second HMGB1 antibody. Using HMV317, nuclear HMGB1 staining is lacking in maturing cells of the spermatogenesis in the testis (A), decreases from the basal cell layers to the top layers in squamous epithelium of the tonsil (B), is variable and often low in proximal tubuli of the kidney (C), and is lower in acinar than in basal cells of the prostate (D). Using clone EPR3507, a comparable staining was seen in the testis (E), tonsil (F), kidney (G), and the prostate (H). Supplementary Figure S2: Further samples of HMGB1 immunostaining observed in different tumor types. (A–E): Tumors with complete loss of HMGB1 expression, including (A) an endometrioid carcinoma of the ovary, (B) clear cell renal cell carcinoma, (C) clear cell carcinoma of the ovary, (D) adenocarcinoma of the colon, and (E) lobular carcinoma of the breast. (F–J): HMGB1-positive tumors, including (F) a breast cancer of no special type, (G) squamous cell carcinoma of the anal canal, (H) neurofibroma, (I) pancreatic adenocarcinoma, and (J) a muscle-invasive urinary bladder cancer. Supplementary Figure S3: Examples of HMGB1 immunostaining scores. (A) Score 0: Absence of HMGB1 staining in a clear cell carcinoma of the ovary, (B) Score 1+: weak HMGB1 staining in a case of adenocarcinoma of the pancreas, (C) Score 2+: HMGB1 staining of moderate intensity in an adenocarcinoma of the colon, and (D) Score 3: strong HMGB1 staining in adenocarcinoma of the colon.

Author Contributions

V.C., H.S., K.J., R.S. and G.S.: contributed to conception, design, data collection, data analysis, and manuscript writing; M.C.T., S.B., C.M.v.B., K.M., F.L., D.D., P.C., M.L., M.F., S.D.R., S.K., V.R., E.B., T.S.C., P.L., C.F., T.K., S.M., A.H.M., W.W. and S.S.: participated in pathology data analysis, data interpretation, and collection of samples; R.S., M.K., G.M.-F. and C.H.-M.: data analysis; K.J., R.S. and G.S.: study supervision. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

The usage of archived diagnostic leftover tissues for manufacturing of TMAs and their analysis for research purposes, as well as patient data analysis, is permitted in accordance with local laws (HmbKHG, §12,1) and has been approved by the local ethics committee (Ethics commission Hamburg, WF-049/09, 25 January 2010). All work has been carried out in compliance with the Helsinki Declaration.

Informed Consent Statement

Patient consent was waived due to local laws (HmbKHG, §12,1) that permit research with anonymized diagnostic leftover tissue samples.

Data Availability Statement

The original contributions presented in the study are included in the article/Supplementary Material, further inquiries can be directed to the corresponding author.

Acknowledgments

We are grateful to Melanie Steurer, Laura Behm, Inge Brandt, and Sünje Seekamp for excellent technical assistance.

Conflicts of Interest

The recombinant rabbit monoclonal antibody, clone HMV317, was provided by MS Validated Antibodies GmbH, Hamburg, Germany (owned by a family member of GS).

References

  1. Ko, Y.B.; Kim, B.R.; Nam, S.L.; Yang, J.B.; Park, S.Y.; Rho, S.B. High-mobility group box 1 (HMGB1) protein regulates tumor-associated cell migration through the interaction with BTB domain. Cell Signal. 2014, 26, 777–783. [Google Scholar] [CrossRef]
  2. Zhu, X.; Messer, J.S.; Wang, Y.; Lin, F.; Cham, C.M.; Chang, J.; Billiar, T.R.; Lotze, M.T.; Boone, D.L.; Chang, E.B. Cytosolic HMGB1 controls the cellular autophagy/apoptosis checkpoint during inflammation. J. Clin. Investig. 2015, 125, 1098–1110. [Google Scholar] [CrossRef]
  3. Chen, R.; Zou, J.; Zhong, X.; Li, J.; Kang, R.; Tang, D. HMGB1 in the interplay between autophagy and apoptosis in cancer. Cancer Lett. 2024, 581, 216494. [Google Scholar] [CrossRef]
  4. Xue, J.; Suarez, J.S.; Minaai, M.; Li, S.; Gaudino, G.; Pass, H.I.; Carbone, M.; Yang, H. HMGB1 as a therapeutic target in disease. J. Cell Physiol. 2021, 236, 3406–3419. [Google Scholar] [CrossRef] [PubMed]
  5. Jube, S.; Rivera, Z.S.; Bianchi, M.E.; Powers, A.; Wang, E.; Pagano, I.; Pass, H.I.; Gaudino, G.; Carbone, M.; Yang, H. Cancer cell secretion of the DAMP protein HMGB1 supports progression in malignant mesothelioma. Cancer Res. 2012, 72, 3290–3301. [Google Scholar] [CrossRef]
  6. Wang, Z.; Wang, X.; Li, J.; Yang, C.; Xing, Z.; Chen, R.; Xu, F. HMGB1 knockdown effectively inhibits the progression of rectal cancer by suppressing HMGB1 expression and promoting apoptosis of rectal cancer cells. Mol. Med. Rep. 2016, 14, 1026–1032. [Google Scholar] [CrossRef] [PubMed]
  7. Yang, Y.; Yang, L.; Jiang, S.; Yang, T.; Lan, J.; Lei, Y.; Tan, H.; Pan, K. HMGB1 mediates lipopolysaccharide-induced inflammation via interacting with GPX4 in colon cancer cells. Cancer Cell Int. 2020, 20, 205. [Google Scholar] [CrossRef]
  8. Li, P.; Ren, K.; Liang, Y.Y.; Liu, J.K.; Liang, Z.W.; Zhang, Y.F. Aloin promotes cell apoptosis by targeting HMGB1-TLR4-ERK axis in human melanoma cells. EXCLI J. 2020, 19, 641–651. [Google Scholar]
  9. De Azevedo, J.; Mourtada, J.; Bour, C.; Devignot, V.; Schultz, P.; Borel, C.; Pencreach, E.; Mellitzer, G.; Gaiddon, C.; Jung, A.C. The EXTREME Regimen Associating Cetuximab and Cisplatin Favors Head and Neck Cancer Cell Death and Immunogenicity with the Induction of an Anti-Cancer Immune Response. Cells 2022, 11, 2866. [Google Scholar] [CrossRef]
  10. Dong, H.; Zhang, L.; Liu, S. Targeting HMGB1: An available Therapeutic Strategy for Breast Cancer Therapy. Int. J. Biol. Sci. 2022, 18, 3421–3434. [Google Scholar] [CrossRef]
  11. Krysko, D.V.; Garg, A.D.; Kaczmarek, A.; Krysko, O.; Agostinis, P.; Vandenabeele, P. Immunogenic cell death and DAMPs in cancer therapy. Nat. Rev. Cancer 2012, 12, 860–875. [Google Scholar] [CrossRef]
  12. Lee, S.A.; Kwak, M.S.; Kim, S.; Shin, J.S. The role of high mobility group box 1 in innate immunity. Yonsei Med. J. 2014, 55, 1165–1176. [Google Scholar] [CrossRef]
  13. Kang, R.; Zhang, Q.; Zeh, H.J., 3rd; Lotze, M.T.; Tang, D. HMGB1 in cancer: Good, bad, or both? Clin. Cancer Res. 2013, 19, 4046–4057. [Google Scholar] [CrossRef]
  14. Wang, L.L.; Meng, Q.H.; Jiao, Y.; Xu, J.Y.; Ge, C.M.; Zhou, J.Y.; Rosen, E.M.; Wang, H.C.; Fan, S.J. High-mobility group boxes mediate cell proliferation and radiosensitivity via retinoblastoma-interaction-dependent and -independent mechanisms. Cancer Biother. Radiopharm. 2012, 27, 329–335. [Google Scholar] [CrossRef]
  15. Shi, Q.; Wang, Y.; Dong, W.; Song, E.; Song, Y. Polychlorinated biphenyl quinone-induced signaling transition from autophagy to apoptosis is regulated by HMGB1 and p53 in human hepatoma HepG2 cells. Toxicol. Lett. 2019, 306, 25–34. [Google Scholar] [CrossRef]
  16. Liang, W.J.; Yang, H.W.; Liu, H.N.; Qian, W.; Chen, X.L. HMGB1 upregulates NF-kB by inhibiting IKB-alpha and associates with diabetic retinopathy. Life Sci. 2020, 241, 117146. [Google Scholar] [CrossRef]
  17. Giavara, S.; Kosmidou, E.; Hande, M.P.; Bianchi, M.E.; Morgan, A.; d’Adda di Fagagna, F.; Jackson, S.P. Yeast Nhp6A/B and mammalian Hmgb1 facilitate the maintenance of genome stability. Curr. Biol. 2005, 15, 68–72. [Google Scholar] [CrossRef]
  18. Celona, B.; Weiner, A.; Di Felice, F.; Mancuso, F.M.; Cesarini, E.; Rossi, R.L.; Gregory, L.; Baban, D.; Rossetti, G.; Grianti, P.; et al. Substantial histone reduction modulates genomewide nucleosomal occupancy and global transcriptional output. PLoS Biol. 2011, 9, e1001086. [Google Scholar] [CrossRef]
  19. Uhlen, M.; Bandrowski, A.; Carr, S.; Edwards, A.; Ellenberg, J.; Lundberg, E.; Rimm, D.L.; Rodriguez, H.; Hiltke, T.; Snyder, M.; et al. A proposal for validation of antibodies. Nat. Methods 2016, 13, 823–827. [Google Scholar] [CrossRef]
  20. Simon, R.; Mirlacher, M.; Sauter, G. Immunohistochemical analysis of tissue microarrays. Methods Mol. Biol. 2010, 664, 113–126. [Google Scholar] [CrossRef]
  21. Hoos, A.; Cordon-Cardo, C. Tissue microarray profiling of cancer specimens and cell lines: Opportunities and limitations. Lab. Invest. 2001, 81, 1331–1338. [Google Scholar] [CrossRef]
  22. Camp, R.L.; Chung, G.G.; Rimm, D.L. Automated subcellular localization and quantification of protein expression in tissue microarrays. Nat. Med. 2002, 8, 1323–1327. [Google Scholar] [CrossRef]
  23. Zhang, C.C.; Gdynia, G.; Ehemann, V.; Roth, W. The HMGB1 protein sensitizes colon carcinoma cells to cell death triggered by pro-apoptotic agents. Int. J. Oncol. 2015, 46, 667–676. [Google Scholar] [CrossRef]
  24. Minamoto, T.; Nakayama, K.; Nakamura, K.; Katagiri, H.; Sultana, R.; Ishibashi, T.; Ishikawa, M.; Yamashita, H.; Sanuki, K.; Iida, K.; et al. Loss of beclin 1 expression in ovarian cancer: A potential biomarker for predicting unfavorable outcomes. Oncol. Lett. 2018, 15, 1170–1176. [Google Scholar] [CrossRef]
  25. Kang, R.; Xie, Y.; Zhang, Q.; Hou, W.; Jiang, Q.; Zhu, S.; Liu, J.; Zeng, D.; Wang, H.; Bartlett, D.L.; et al. Intracellular HMGB1 as a novel tumor suppressor of pancreatic cancer. Cell Res. 2017, 27, 916–932. [Google Scholar] [CrossRef]
  26. Luan, X.; Ma, C.; Wang, P.; Lou, F. HMGB1 is negatively correlated with the development of endometrial carcinoma and prevents cancer cell invasion and metastasis by inhibiting the process of epithelial-to-mesenchymal transition. OncoTargets Ther. 2017, 10, 1389–1402. [Google Scholar] [CrossRef]
  27. Lu, H.; Zhu, M.; Qu, L.; Shao, H.; Zhang, R.; Li, Y. Oncogenic Role of HMGB1 as An Alarming in Robust Prediction of Immunotherapy Response in Colorectal Cancer. Cancers 2022, 14, 4875. [Google Scholar] [CrossRef]
  28. Yu, S.; Qian, L.; Ma, J. Genetic alterations, RNA expression profiling and DNA methylation of HMGB1 in malignancies. J. Cell Mol. Med. 2022, 26, 4322–4332. [Google Scholar] [CrossRef]
  29. Zhang, C.; Ge, S.; Hu, C.; Yang, N.; Zhang, J. MiRNA-218, a new regulator of HMGB1, suppresses cell migration and invasion in non-small cell lung cancer. Acta Biochim. Biophys. Sin. 2013, 45, 1055–1061. [Google Scholar] [CrossRef]
  30. Guerriero, J.L.; Ditsworth, D.; Catanzaro, J.M.; Sabino, G.; Furie, M.B.; Kew, R.R.; Crawford, H.C.; Zong, W.X. DNA alkylating therapy induces tumor regression through an HMGB1-mediated activation of innate immunity. J. Immunol. 2011, 186, 3517–3526. [Google Scholar] [CrossRef]
  31. Shrivastava, S.; Mansure, J.J.; Almajed, W.; Cury, F.; Ferbeyre, G.; Popovic, M.; Seuntjens, J.; Kassouf, W. The Role of HMGB1 in Radioresistance of Bladder Cancer. Mol. Cancer Ther. 2016, 15, 471–479. [Google Scholar] [CrossRef]
  32. Zhang, X.; Yang, X.; Zhu, S.; Li, Q.; Zou, N. Radiosensitization of esophageal carcinoma cells by knockdown of HMGB1 expression. Oncol. Rep. 2019, 41, 1960–1970. [Google Scholar] [CrossRef]
  33. Yamazaki, T.; Hannani, D.; Poirier-Colame, V.; Ladoire, S.; Locher, C.; Sistigu, A.; Prada, N.; Adjemian, S.; Catani, J.P.; Freudenberg, M.; et al. Defective immunogenic cell death of HMGB1-deficient tumors: Compensatory therapy with TLR4 agonists. Cell Death Differ. 2014, 21, 69–78. [Google Scholar] [CrossRef]
  34. Bottger, F.; Radonic, T.; Bahce, I.; Monkhorst, K.; Piersma, S.R.; Pham, T.V.; Dingemans, A.C.; Hillen, L.M.; Santarpia, M.; Giovannetti, E.; et al. Identification of protein biomarkers for prediction of response to platinum-based treatment regimens in patients with non-small cell lung cancer. Mol. Oncol. 2024, 18, 1417–1436. [Google Scholar] [CrossRef]
  35. Li, P.; Xu, M.; Cai, H.; Thapa, N.; He, C.; Song, Z. The effect of HMGB1 on the clinicopathological and prognostic features of cervical cancer. Biosci. Rep. 2019, 39, BSR20181016. [Google Scholar] [CrossRef]
  36. Zhang, X.; Zou, N.; Deng, W.; Song, C.; Yan, K.; Shen, W.; Zhu, S. HMGB1 induces radioresistance through PI3K/AKT/ATM pathway in esophageal squamous cell carcinoma. Mol. Biol. Rep. 2022, 49, 11933–11945. [Google Scholar] [CrossRef]
  37. Su, C.; Jia, S.; Ma, Z.; Zhang, H.; Wei, L.; Liu, H. HMGB1 Promotes Lymphangiogenesis through the Activation of RAGE on M2 Macrophages in Laryngeal Squamous Cell Carcinoma. Dis. Markers 2022, 2022, 4487435. [Google Scholar] [CrossRef]
  38. Huang, C.; Huang, Z.; Zhao, X.; Wang, Y.; Zhao, H.; Zhong, Z.; Wang, L. Overexpression of high mobility group box 1 contributes to progressive clinicopathological features and poor prognosis of human bladder urothelial carcinoma. OncoTargets Ther. 2018, 11, 2111–2120. [Google Scholar] [CrossRef]
  39. Taguchi, R.; Yamaguchi-Tanaka, M.; Takagi, K.; Sato, A.; Miki, Y.; Miyashita, M.; Suzuki, T. Clinicopathological Significance and Prognostic Role of High Mobility Group Box 1 (HMGB1), Toll-Like Receptor (TLR) 2 and TLR4 in Breast Cancer. Acta Histochem. Cytochem. 2024, 57, 75–83. [Google Scholar] [CrossRef]
  40. Dong, J.; Li, J.; Liu, S.; Feng, X.; Chen, S.; Zhou, Z.; Chen, Y.; Zhang, X. Prognostic potential of an immune score based on the density of CD8(+) T cells, CD20(+) B cells, and CD33(+)/p-STAT1(+) double-positive cells and HMGB1 expression within cancer nests in stage IIIA gastric cancer patients. Chin. J. Cancer Res. 2016, 28, 543–552. [Google Scholar] [CrossRef]
  41. Suren, D.; Arda Gokay, A.; Sayiner, A. High Mobility Group Box 1 (HMGB1) expression in gastric adenocarcinomas. J. BUON 2018, 23, 422–427. [Google Scholar] [PubMed]
  42. Cao, X.P.; Cao, Y.; Zhao, H.; Yin, J.; Hou, P. HMGA1 promoting gastric cancer oncogenic and glycolytic phenotypes by regulating c-myc expression. Biochem. Biophys. Res. Commun. 2019, 516, 457–465. [Google Scholar] [CrossRef] [PubMed]
Diagnostics 15 01974 g001
Figure 1. HMGB1 immunostaining of normal tissues. The panels show a strong nuclear staining in lymphocytes, epithelial and stroma cells of the tonsil (A), as well as in epithelial and stromal cells of the colon (B) and the endometrium (C) while the HMGB1 staining intensity decreased from the basal towards the superficial cell layers in squamous epithelium of the oral mucosa (D). A less intense nuclear staining was seen in proximal tubules of the kidney (E) and in acinar cells of the prostate gland (F). HMGB1 staining was absent or only weak in medullary cells of the adrenal gland (G), and in maturing cells of the spermatogenesis in the testis (H).
Figure 1. HMGB1 immunostaining of normal tissues. The panels show a strong nuclear staining in lymphocytes, epithelial and stroma cells of the tonsil (A), as well as in epithelial and stromal cells of the colon (B) and the endometrium (C) while the HMGB1 staining intensity decreased from the basal towards the superficial cell layers in squamous epithelium of the oral mucosa (D). A less intense nuclear staining was seen in proximal tubules of the kidney (E) and in acinar cells of the prostate gland (F). HMGB1 staining was absent or only weak in medullary cells of the adrenal gland (G), and in maturing cells of the spermatogenesis in the testis (H).
Diagnostics 15 01974 g001
Diagnostics 15 01974 g002
Figure 2. HMGB1 immunostaining in cancer. The panels show a complete absence of nuclear HMGB1 staining in all tumor cells but retained staining in non-neoplastic cells of a phaeochromocytoma (A), a seminoma (B), a gastrointestinal stroma tumor (C), a Hodgkin’s lymphoma (D), a clear cell renal cell carcinoma (E), and a prostatic adenocarcinoma (F). Examples of HMGB1-positive tumors include a serous adenocarcinoma of the ovary (G) and an adenocarcinoma of the colon (H).
Figure 2. HMGB1 immunostaining in cancer. The panels show a complete absence of nuclear HMGB1 staining in all tumor cells but retained staining in non-neoplastic cells of a phaeochromocytoma (A), a seminoma (B), a gastrointestinal stroma tumor (C), a Hodgkin’s lymphoma (D), a clear cell renal cell carcinoma (E), and a prostatic adenocarcinoma (F). Examples of HMGB1-positive tumors include a serous adenocarcinoma of the ovary (G) and an adenocarcinoma of the colon (H).
Diagnostics 15 01974 g002
Diagnostics 15 01974 g003
Figure 3. Ranking of human tumor types according to the frequency of HMGB1 expression loss.
Figure 3. Ranking of human tumor types according to the frequency of HMGB1 expression loss.
Diagnostics 15 01974 g003
Table 1. HMGB1 immunostaining in human tumors. int. = interpretable.
Table 1. HMGB1 immunostaining in human tumors. int. = interpretable.
On TMA (n)HMGB1 Immunostaining
Tumor Entityint. (n)0 (%)1+ (%)2+ (%)3+ (%)
Tumors of the skinBasal cell carcinoma of the skin41260.00.00.0100.0
Squamous cell carcinoma of the skin95813.711.150.634.6
Malignant melanoma19195.310.547.436.8
Malignant melanoma lymph node metastasis86633.26.320.669.8
Merkel cell carcinoma220.050.050.00.0
Tumors of the head and neckSquamous cell carcinoma of the larynx109724.216.722.256.9
Squamous cell carcinoma of the pharynx60470.014.931.953.2
Oral squamous cell carcinoma (floor of the mouth)130953.213.735.847.4
Pleomorphic adenoma of the parotid gland50210.09.54.885.7
Warthin tumor of the parotid gland49320.00.015.684.4
Basal cell adenoma of the salivary gland1590.00.022.277.8
Tumors of the lung, pleura, and thymusAdenocarcinoma of the lung1961452.15.538.653.8
Squamous cell carcinoma of the lung80551.816.430.950.9
Mesothelioma, epithelioid402810.725.025.039.3
Mesothelioma, biphasic29150.026.733.340.0
Thymoma29180.011.111.177.8
Lung, neuroendocrine tumor (NET)292711.13.718.566.7
Tumors of the
female genital tract		Squamous cell carcinoma of the vagina30229.19.131.850.0
Squamous cell carcinoma of the vulva107891.116.924.757.3
Squamous cell carcinoma of the cervix88781.320.539.738.5
Adenocarcinoma of the cervix23220.04.513.681.8
Endometrioid endometrial carcinoma2882610.810.039.849.4
Endometrial serous carcinoma36310.03.232.364.5
Carcinosarcoma of the uterus574910.214.330.644.9
Endometrial carcinoma, high grade, G313137.723.138.530.8
Endometrial clear cell carcinoma9812.537.525.025.0
Endometrioid carcinoma of the ovary93710.012.732.454.9
Serous carcinoma of the ovary5304420.713.133.952.3
Mucinous carcinoma of the ovary75520.07.721.271.2
Clear cell carcinoma of the ovary51419.817.134.139.0
Carcinosarcoma of the ovary47370.02.724.373.0
Granulosa cell tumor of the ovary44350.08.614.377.1
Leydig cell tumor of the ovary430.066.70.033.3
Sertoli cell tumor of the ovary110.00.0100.00.0
Sertoli Leydig cell tumor of the ovary330.00.00.0100.0
Steroid cell tumor of the ovary3333.333.333.30.0
Brenner tumor32280.025.07.167.9
Tumors of the breastInvasive breast carcinoma of no special type4994026.713.737.342.3
Lobular carcinoma of the breast1501022.013.740.244.1
Medullary carcinoma of the breast870.00.071.428.6
Tubular carcinoma of the breast210.00.00.0100.0
Mucinous carcinoma of the breast7714.314.328.642.9
Tumors of the
digestive system		Adenomatous polyp, low-grade dysplasia50340.00.02.997.1
Adenomatous polyp, high-grade dysplasia50450.02.22.295.6
Adenocarcinoma of the colon248320970.23.920.675.3
Gastric adenocarcinoma, diffuse type2151401.410.728.659.3
Gastric adenocarcinoma, intestinal type2151550.611.026.561.9
Gastric adenocarcinoma, mixed type62450.020.033.346.7
Adenocarcinoma of the esophagus83420.00.021.478.6
Squamous cell carcinoma of the esophagus76385.313.221.160.5
Squamous cell carcinoma of the anal canal91663.010.625.860.6
Cholangiocarcinoma58434.711.632.651.2
Gallbladder adenocarcinoma51442.34.531.861.4
Gallbladder Klatskin tumor42350.08.614.377.1
Hepatocellular carcinoma3122807.114.333.944.6
Ductal adenocarcinoma of the pancreas6594650.67.534.257.6
Pancreatic/Ampullary adenocarcinoma98681.510.336.851.5
Acinar cell carcinoma of the pancreas18156.713.333.346.7
Gastrointestinal stromal tumor (GIST)625628.632.126.812.5
Appendix, neuroendocrine tumor (NET)25100.020.030.050.0
Colorectal, neuroendocrine tumor (NET)1290.011.133.355.6
Ileum, neuroendocrine tumor (NET)53462.26.528.363.0
Pancreas, neuroendocrine tumor (NET)101821.24.931.762.2
Colorectal, neuroendocrine carcinoma (NEC)14120.08.333.358.3
Ileum, neuroendocrine carcinoma (NEC)870.028.614.357.1
Gallbladder, neuroendocrine carcinoma (NEC)440.00.050.050.0
Pancreas, neuroendocrine carcinoma (NEC)14100.010.050.040.0
Tumors of the
urinary system		Non-invasive papillary urothelial ca., pTa G2 low grade87780.01.33.894.9
Non-invasive papillary urothelial ca., pTa G2 high grade80680.00.04.495.6
Non-invasive papillary urothelial car., pTa G31261031.01.914.682.5
Urothelial carcinoma, pT2–4 G37355622.19.329.259.4
Squamous cell carcinoma of the bladder22190.021.110.568.4
Small cell neuroendocrine carcinoma of the bladder550.00.00.0100.0
Sarcomatoid urothelial carcinoma25166.318.831.343.8
Urothelial carcinoma of the kidney pelvis62504.00.016.080.0
Clear cell renal cell carcinoma1287105012.77.721.957.7
Papillary renal cell carcinoma3682858.49.127.754.7
Clear cell (tubulo) papillary renal cell carcinoma26180.05.65.688.9
Chromophobe renal cell carcinoma17014216.214.128.241.5
Oncocytoma of the kidney25719510.311.833.344.6
Tumors of the male genital
organs		Adenocarcinoma of the prostate, Gleason 3+3834916.332.732.718.4
Adenocarcinoma of the prostate, Gleason 4+480568.932.137.521.4
Adenocarcinoma of the prostate, Gleason 5+5856716.420.941.820.9
Adenocarcinoma of the prostate (recurrence)2582229.518.938.333.3
Small cell neuroendocrine carcinoma of the prostate210.00.00.0100.0
Seminoma68257972.418.38.60.7
Embryonal carcinoma of the testis54427.147.638.17.1
Leydig cell tumor of the testis312416.733.333.316.7
Sertoli cell tumor of the testis220.00.00.0100.0
Sex cord stromal tumor of the testis110.0100.00.00.0
Spermatocytic tumor of the testis110.0100.00.00.0
Yolk sac tumor533915.435.930.817.9
Teratoma53350.020.05.774.3
Squamous cell carcinoma of the penis92735.58.241.145.2
Tumors of
endocrine organs		Adenoma of the thyroid gland63490.02.020.477.6
Papillary thyroid carcinoma3412690.41.913.884.0
Follicular thyroid carcinoma109560.03.623.273.2
Medullary thyroid carcinoma57450.013.331.155.6
Parathyroid gland adenoma43270.00.00.0100.0
Anaplastic thyroid carcinoma19140.028.650.021.4
Adrenal cortical adenoma483710.818.924.345.9
Adrenal cortical carcinoma272425.033.325.016.7
Pheochromocytoma513090.010.00.00.0
Tumors of
hematopoietic and lymphoid tissues		Hodgkin’s lymphoma1034825.031.341.72.1
Small lymphocytic lymphoma, B-cell type (B-SLL/B-CLL)50450.00.06.793.3
Diffuse large B-cell lymphoma (DLBCL)1131031.015.528.255.3
Follicular lymphoma88830.00.02.497.6
T-cell non-Hodgkin’s lymphoma25210.023.823.852.4
Mantle cell lymphoma18170.00.00.0100.0
Marginal zone lymphoma16150.06.70.093.3
Diffuse large B-cell lymphoma (DLBCL) in the testis16150.033.36.760.0
Burkitt lymphoma530.00.00.0100.0
Tumors of soft tissue and boneGranular cell tumor23156.70.020.073.3
Leiomyoma50368.32.816.772.2
Leiomyosarcoma94745.417.623.054.1
Liposarcoma96736.811.021.960.3
Malignant peripheral nerve sheath tumor (MPNST)15120.016.725.058.3
Myofibrosarcoma26258.016.048.028.0
Angiosarcoma42273.77.422.266.7
Angiomyolipoma91763.96.67.981.6
Dermatofibrosarcoma protuberans21120.08.325.066.7
Ganglioneuroma14110.00.09.190.9
Kaposi sarcoma840.00.025.075.0
Neurofibroma117867.00.07.086.0
Sarcoma, not otherwise specified (NOS)74639.512.719.058.7
Paraganglioma413212.56.334.446.9
Ewing sarcoma23128.38.325.058.3
Rhabdomyosarcoma7616.70.016.766.7
Schwannoma1221022.90.03.993.1
Synovial sarcoma1280.00.037.562.5
Osteosarcoma19128.38.316.766.7
Chondrosarcoma15714.328.60.057.1
Rhabdoid tumor540.00.025.075.0
Solitary fibrous tumor17175.90.023.570.6
Table 2. HMGB1 immunostaining and tumor phenotype.
Table 2. HMGB1 immunostaining and tumor phenotype.
Tumor EntityPathological and Molecular Parameters HMGB1 Immunostaining
nNegative (%)Weak (%)Moderate (%)Strong (%)pOR (95%CI)
Invasive breast carcinoma of no special typepT11432.813.333.650.30.00380.19 (0.04–0.95)
pT217210.510.540.738.4
pT3–4339.130.330.330.3
G1110054.545.50.37491.48 (0.43–5.11)
G21928.312.53742.2
G31496.715.434.943
pN01745.712.136.2460.23220.25 (0.02–3.02)
pN1919.916.534.139.6
pN2397.712.843.635.9
pN3147.128.65014.3
Clear cell renal cell carcinomaISUP 123312.46.917.663.1<0.00010.16 (0.07–0.37)
ISUP 23488.37.520.164.1
ISUP 322921.81027.540.6
ISUP 46225.816.137.121
Fuhrman 1547.41.916.774.1<0.00010.11 (0.03–0.38)
Fuhrman 26019.2719.164.7
Fuhrman 325721.49.726.842
Fuhrman 47622.415.836.825
Thoenes 130510.25.618.765.6<0.00010.22 (0.11–0.44)
Thoenes 241917.210.526.545.8
Thoenes 38223.215.92832.9
UICC 127510.25.521.562.9<0.00010.14 (0.07–0.31)
UICC 23212.59.431.346.9
UICC 37915.220.325.339.2
UICC 45832.815.522.429.3
pT15957.7519.268.1<0.00010.18 (0.11–0.27)
pT211314.211.529.245.1
pT3–428523.9132637.2
pN014716.39.526.547.60.01080.15 (0.03–0.61)
pN+2330.41343.513
pM09312.97.530.149.50.04130.36 (0.15–0.85)
pM+7824.415.426.933.3
Urothelial bladder carcinomapTa G2 low7801.33.894.90.007864.83 (5.83–720.42)
pTa G2 high68004.495.6
pTa G3811.22.518.577.8
pT21181.712.727.158.50.21520.54 (0.11–2.75)
pT32062.99.733.553.9
pT410106.933.759.4
G222013.618.268.20.3532 *1.15 (0.32–4.15)
G340329.732.555.8
pN02331.79.934.354.10.6253 *0.78 (0.19–3.24)
pN+1642.48.529.359.8
L01661.21234.352.40.2492 *0.53 (0.09–2.96)
L11552.67.130.360
V02341.710.335.552.60.4817 *0.73 (0.13–4.12)
V1752.79.326.761.3
Endometrioid endometrial carcinomapT110306.848.544.70.16930.91 (0.21–3.9)
pT2244.216.72554.2
pT3–43508.64051.4
pN04906.142.9510.15880.27 (0.05–1.44)
pN+293.413.851.731
Serous carcinoma of
the ovary		pT1293.424.127.644.80.18412.24 (0.82–6.1)
pT241019.531.748.8
pT3244012.336.551.2
pN0781.324.435.938.50.02132.76 (1.3–5.86)
pN+156012.831.455.8
Adenocarcinoma of the colorectumpT17601.314.584.20.10170.22 (0.03–1.73)
pT238902.818.378.9
pT311270.44.421.274
pT43970522.272.8
pN010230.43.422.373.90.01370.7 (0.45–1.09)
pN+9610519.375.8
V014240.24.420.874.60.6021.04 (0.11–9.68)
V15300.23.419.177.4
L06260.24.518.776.70.46450.96 (0.12–8.09)
L113320.24.121.874
Left side11110.22.22176.70.01290.66 (0.07–6.13)
Right side3970.35.518.475.8
MMR-deficient7305.530.164.40.01672.87 (0.96–8.58)
MMR-proficient10230.12.416.880.6
RAS mutation3480.6220.776.70.15660.96 (0.36–2.55)
RAS wildtype45302.21780.8
BRAF V600E mutation210038.161.90.05662.93 (1.08–7.94)
BRAF wildtype1260416.779.4
Adenocarcinoma of the pancreaspT180025750.65710.48 (0.07–3.09)
pT249010.226.563.3
pT32850.75.63459.6
pT4195.35.336.852.6
G1110027.372.70.50050.54 (0.13–2.22)
G22561.25.932.460.5
G3780937.253.8
pN0710735.257.70.63891.25 (0.44–3.6)
pN+28915.932.260.9
Adenocarcinoma of the stomachpT1–2452.215.635.646.70.79252.19 (0.13–36.73)
pT31041.923.133.741.3
pT4101114.937.646.5
pN0631.614.331.752.40.55530.81 (0.08–7.98)
pN+1871.619.336.942.2
MMR-deficient 30013.343.343.30.58881.02 (0.31–3.35)
MMR-proficient2202.314.53548.2
SeminomaspT130769.719.99.80.70.69613.59 (0.33–39.49)
pT21207021.77.50.8
pT34858.32514.62.1
Haemangioinvasion negative3946721.810.70.50.17067.95 (1.08–58.49)
Positive4870.818.86.34.2
Lymphangioinvasion negative34167.221.710.90.30.15299.47 (0.99–90.33)
Positive10568.6208.62.9
Infiltration of the spermatic cord negative37070209.20.80.50832.62 (0.26–26.27)
Positive5263.519.215.41.9
Infiltration Rete Testis negative21169.219.910.40.50.84551.56 (0.14–16.91)
Positive2417119.98.30.8
Squamous cell carcinomas of different sites **pT11701.210.63058.20.14280.18 (0.04–0.93)
pT21923.615.133.947.4
pT31113.613.528.854.1
pT41085.620.424.150
G1219.5061.928.60.0017.51 (1.14–49.43)
G22764.317.826.151.8
G31772.311.334.552
pN02304.313.52755.20.34862.26 (0.68–7.49)
pN+2181.815.130.752.3
* only in pT2–4 urothelial bladder carcinomas, ** oral, pharynx, larynx, esophagus, cervix, vagina, vulva, penis, anal, and lung; abbreviation: pT: pathological tumor stage, G: grade, pN: pathological lymph node status, pM: pathological status of distant metastasis, R: resection margin status, V: venous invasion, L: lymphatic invasion, MMR: mismatch repair, ISUP: International Society of Urological Pathology, UICC: Union for International Cancer Control, OR: odds ratios are provided for the lowest vs. the highest parameter.
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MDPI and ACS Style

Chirico, V.; Sharifi, H.; Tsourlakis, M.C.; Büyücek, S.; Bargen, C.M.v.; Möller, K.; Lutz, F.; Dum, D.; Kluth, M.; Hube-Magg, C.; et al. HMGB1 Deficiency Occurs in a Broad Range of Human Cancers and Is Often Associated with Unfavorable Tumor Phenotype. Diagnostics 2025, 15, 1974. https://doi.org/10.3390/diagnostics15151974

AMA Style

Chirico V, Sharifi H, Tsourlakis MC, Büyücek S, Bargen CMv, Möller K, Lutz F, Dum D, Kluth M, Hube-Magg C, et al. HMGB1 Deficiency Occurs in a Broad Range of Human Cancers and Is Often Associated with Unfavorable Tumor Phenotype. Diagnostics. 2025; 15(15):1974. https://doi.org/10.3390/diagnostics15151974

Chicago/Turabian Style

Chirico, Viktoria, Hena Sharifi, Maria Christina Tsourlakis, Seyma Büyücek, Clara Marie von Bargen, Katharina Möller, Florian Lutz, David Dum, Martina Kluth, Claudia Hube-Magg, and et al. 2025. "HMGB1 Deficiency Occurs in a Broad Range of Human Cancers and Is Often Associated with Unfavorable Tumor Phenotype" Diagnostics 15, no. 15: 1974. https://doi.org/10.3390/diagnostics15151974

APA Style

Chirico, V., Sharifi, H., Tsourlakis, M. C., Büyücek, S., Bargen, C. M. v., Möller, K., Lutz, F., Dum, D., Kluth, M., Hube-Magg, C., Makrypidi-Fraune, G., Caneve, P., Lennartz, M., Freytag, M., Dwertmann Rico, S., Kind, S., Reiswich, V., Burandt, E., Clauditz, T. S., ... Jansen, K. (2025). HMGB1 Deficiency Occurs in a Broad Range of Human Cancers and Is Often Associated with Unfavorable Tumor Phenotype. Diagnostics, 15(15), 1974. https://doi.org/10.3390/diagnostics15151974

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