Hironori Haga | Kyoto University (original) (raw)

Papers by Hironori Haga

Annals of Oncology, Sep 1, 2021

Transplantation, May 1, 2017

Background. The cause of late graft dysfunction has not been elucidated. Although an antibody-med... more Background. The cause of late graft dysfunction has not been elucidated. Although an antibody-mediated reaction is suspected as a potential mechanism, the target antigens have not been clarified. Methods. To clarify the etiology of idiopathic posttransplantation hepatitis (IPTH), we simultaneously examined the presence of antibodies that react with liver tissue (ARLT) by means of indirect immunofluorescence staining, as well as the presence of donor-specific anti-human leukocyte antigen antibodies (HLA-DSA). A subanalysis of the IPTH group was also performed. Within the IPTH group, the correlation between ARLT titer and clinical data were analyzed. Results. In the sera of patients with IPTH (30 patients), ARLT were found at a significantly higher frequency than in patients without IPTH (42 patients; P < 0.001). Moreover, the ARLT titer appeared to be correlated with the severity of hepatitis or hepatic injury. In contrast, the frequency of HLA-DSA was significantly lower in patients with IPTH than in patients without IPTH (P = 0.001). Conclusion. Our findings indicate that ARLT, and not HLA-DSA, profoundly influence the etiology of IPTH.

[Introduction] Non-malignant proliferative lesions in the breast have been implicated in the deve... more [Introduction] Non-malignant proliferative lesions in the breast have been implicated in the development of invasive breast cancer. Previous studies showed that adjacent atypical proliferative lesions and breast cancers shared common genetic alterations, suggesting that these evolved from the same ancestral cell. However, the clonal structure of atypical proliferative lesions and their clonal dynamics during progression to cancer are poorly understood. In this study, we compared genetic profiles of normal ducts, non-malignant proliferative lesions and cancers in the same patients to illustrate the clonal evolution of cancer from a non-malignant epithelial cell. [Methods] Multiple samples were collected from different atypical proliferative lesions within the cancer-borne breast, including invasive cancers, using micro-dissection from formalin-fixed, paraffin-embedded surgical specimens. Somatic mutations and copy number alterations (CNAs) were then evaluated by whole exome sequencing. [Results] A total of 34 samples from 5 premenopausal females carrying estrogen receptor-positive cancers were analyzed, where the samples were obtained from normal ducts (N = 6), non-atypical (N = 1) and atypical (N = 8) proliferative lesions, and invasive (N = 3) and non-invasive (N = 16) cancers. The number of somatic mutations per sample was ranging from 1 to 276 and increased with an elevation of atypical-level. Two cases with bilateral cancers had a pathogenic germline mutation of either BRCA2 and TP53, where no somatic mutations or CNAs were shared by individual proliferative lesions, suggesting multifocal independent cancerous evolutions. By contrast, in the remaining three unilateral cases, no pathogenic germline mutations were detected, but all proliferative lesions, which were separated by a distance of 7-25 mm, shared one or more driver alterations, such as an AKT1 mutation (UID: KU01), concurrent 1q gain and 16q loss (der(1;16)) (UID: KU02), and a GATA3 mutation and der(1;16) (UID: KU03), while harboring private mutations/CNAs of their own. The analysis of phylogenic trees based on the number of shared mutations predicted an early origin of these founder mutations, which frequently predated decades before the onset of cancer. [Conclusions] Early breast cancer development is thought to be shaped by the simultaneous evolution of multiple precancerous clones. It may be multi-focally initiated by a germline mutation, frequently terminated in bilateral cancers. Otherwise, in unilateral cases, cancer clones might be derived from a common ancestral clone, which has acquired a driver founder mutation long before the onset of cancer, and undergo independent evolution, giving rise to multiple proliferative lesions, from which invasive cancer finally evolves. Our findings provide unique insight into the early development of breast cancer. Citation Format: Tomomi Nishimura, Kenichi Yoshida, Yukiko Kawata, Yasuhide Takeuchi, Nobuyuki Kakiuchi, Yusuke Shiozawa, Kosuke Aoki, Masahiro Hirata, Tatsuki R. Kataoka, Takaki Sakurai, Yuichi Shiraishi, Kenichi Chiba, Kengo Takeuchi, Hironori Haga, Satoru Miyano, Masakazu Toi, Seishi Ogawa. Clonal evolution of atypical proliferative lesions into breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3389.

Transplantation, Feb 1, 2007

We retrospectively determined the correlation of results of lymphocyte crossmatch tests by direct... more We retrospectively determined the correlation of results of lymphocyte crossmatch tests by direct complement-dependent cytotoxicity, to the outcomes of 585 consecutive ABO-identical and human leukocyte antigen (HLA)-mismatched living donor liver transplants (LDLTs) (male:female=276:309; median age, 18 years). Crossmatch test results were positive in 14 recipients (2.4%). Patient survival at eight years in the crossmatch-positive group was significantly lower than in the crossmatch-negative group (positive group, 56.3%; negative group, 77.6%; P=0.014). The survival at five years of the crossmatch-positive group was significantly lower than the negative group in both older recipients (&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;or=18 years of age: positive group, 41.7%; negative group, 76.4%; P=0.0065), and female recipients (positive group, 37.5%; negative group, 81.9%; P=3.3x10). We conclude that antidonor antibodies have adverse effects on the clinical outcome of LDLTs, and that being female and/or older aged (&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;or=18 years of age) are risk factors for LDLT.

Liver Transplantation, 2007

Although the effectiveness of rituximab has been reported in ABO blood group (ABO)-incompatible (... more Although the effectiveness of rituximab has been reported in ABO blood group (ABO)-incompatible (ABO-I) organ transplantation, the protocol is not yet established. We studied the impact of the timing of rituximab prophylaxis and the humoral immune response of patients undergoing ABO-I living donor liver transplantation (LDLT), focusing on clinicopathological findings and the B-cell subset. From July 2003 to December 2005, 30 adult patients were treated with hepatic artery infusion (HAI) protocol without splenectomy for ABO-I LDLT. A total of 17 patients were treated only with HAI (no prophylaxis), and the other 13 were treated with rituximab prophylaxis at various times prior to transplantation. For B-cell study of the spleen, another 4 patients undergoing ABO-I LDLT both with HAI after prophylaxis and eventual splenectomy, and 3 patients with ABOcompatible LDLT with splenectomy were enrolled. The mortality of the 30 patients with HAI, without splenectomy, and with/without rituximab prophylaxis was 33% and the main cause of death was sepsis. Peripheral blood B cells were completely depleted, anti-donor blood-type antibody titer was lower, and clinical and pathological antibody-mediated rejection was not observed in patients with prophylaxis earlier than 7 days before transplantation (early prophylaxis). Early rituximab prophylaxis significantly depleted B cells and memory B cells in the spleen but not in lymph nodes. On the other hand, B cells and memory B cells increased and memory B cells became dominant during antibody-mediated rejection. In conclusion, early prophylaxis with rituximab depletes B cells, including memory B cells, in the spleen and is associated with a trend toward lower humoral rejection rates and lower peak immunoglobulin (Ig)G titers in ABO-I LDLT patients.

Molecular and Cellular Biology / Genetics, Jul 1, 2019

Transplantation Proceedings, Apr 1, 2023

Liver Transplantation, 2006

Patient selection criteria of deceased donor liver transplantation for primary biliary cirrhosis ... more Patient selection criteria of deceased donor liver transplantation for primary biliary cirrhosis (PBC) are almost completely established. The aim of this study was to establish selection criteria for both patients and donors of living donor liver transplantation (LDLT) for PBC. We used univariate and multivariate analyses to examine patient and donor characteristics of our first 50 cases of LDLT for PBC to elucidate factors that significantly impacted patient survival or disease recurrence after LDLT in the univariate and/or multivariate analyses. Multivariate analysis demonstrated that the presence of persistent ascites before LDLT, a higher number of human leukocyte antigen (HLA)-A,-B, and-DR mismatches between donor and recipient, and donor age Ն50 years were factors significantly associated with early posttransplant death. Independent risk factors for PBC recurrence after LDLT were a lower number of HLA mismatches between donor and recipient, and a lower average trough level of tacrolimus within 1 year after LDLT. Specifically, the lower the number of HLA-A,-B, and-DR mismatches or the average trough level of tacrolimus within 1 year after LDLT, the higher the possibility of developing a recurrence of PBC. In conclusion, the absence of persistent ascites before LDLT, a lower number of HLA-A,-B, and-DR mismatches between donor and recipient, and a younger donor (Ͻ50 years) are preferred for gaining acceptable survival outcomes for the transplant. However, a lower number of HLA-A,-B, and-DR mismatches between donor and recipient may be a risk factor for PBC recurrence.

Lung Cancer, Sep 1, 2014

Objectives: While novel anti-human epidermal growth factor receptor 2 (HER2) agents have recently... more Objectives: While novel anti-human epidermal growth factor receptor 2 (HER2) agents have recently been developed, no definite criteria have been proposed as indications for the use of these agents in patients with lung cancer. Here, we tested HER2 alterations by using four methods and explored the concordance of these methods to improve our understanding of the accuracy of HER2 testing methods. Materials and methods: We analyzed HER2 protein expression by immunohistochemistry (IHC) and HER2 amplification by fluorescence in situ hybridization (FISH) and dual in situ hybridization (DISH) by using a tissue microarray comprising lung adenocarcinoma specimens from 243 consecutive patients. The presence of mutations in the EGFR, KRAS, and HER2 genes were also determined. Results: Positive IHC (score 3+) was observed in six cases (2.5%). Amplification of HER2 was observed in five cases (2.1%) by FISH and in nine cases (3.7%) by DISH. HER2 expression by IHC and gene amplification by FISH were significantly associated (P < 0.001). The overall concordance between FISH and DISH by amplification status was 96.7% (P < 0.001). One hundred nine tumors (49.9%) had EGFR mutations, 25 (11.2%) had KRAS mutations, and six (2.7%) had HER2 mutations. All of these mutations were mutually exclusive. Cases having HER2 mutations were positively correlated with cases having HER2 amplification (P < 0.001). Two of six cases with HER2 mutations showed amplifications by FISH and DISH tests. Conclusion: HER2 protein overexpression, gene amplification, and gene mutations appeared to be uncommon in lung adenocarcinoma. Cases with HER2 mutations tended to show HER2 gene amplification. The results indicated that HER2 gene amplification and mutations should be tested to determine whether patients are eligible for administration of new anti-HER2 agents. In addition, DISH was better than FISH for detection of cases with HER2 amplification.

Transplantation, Feb 27, 2009

Cancer Research, Jun 15, 2022

[Introduction] Previous studies suggested that breast cancers (BCs) and some adjacent atypical pr... more [Introduction] Previous studies suggested that breast cancers (BCs) and some adjacent atypical proliferative lesions evolved from the same ancestral cell. However, the clonal structure of normal epithelial cells and their clonal history during evolution to cancer are poorly understood. In this study, we analyzed genetic profiles of normal epithelia and proliferative lesions in the cancer-borne breast to illustrate the clonal evolution of cancer from a normal epithelial cell. [Methods] Single cell-derived normal epithelial organoids (n=47) were established from breast milk of 4 healthy women aged 22-36 and surgical specimens of 15 BC patients aged 29-83 to evaluate somatic mutation rate. Multiple normal lobules or proliferative lesions together with cancer cells were collected using laser-capture micro-dissection (LCM) from fresh frozen (n=3) or formalin-fixed paraffin-embedded (n=6) surgical specimens in 9 premenopausal BC patients. Somatic mutations and copy number alterations were evaluated using whole-genome sequencing. [Results] On the basis of single cell-derived mammary gland-derived organoids, somatic mutations increased in mammary gland cells at a constant rate of 19.4/genome/year before menopause, which then decreased to 6.9/genome/year after menopause. Parity was negatively associated with mutation number (-49.3 per life birth). In total, we analyzed 113 LCM samples, including those from 66 normal lobules, 28 proliferative lesions, and 14 non-invasive and 5 invasive cancer samples. Phylogenetic analysis showed that most of the neighboring normal lobules (53/66) from 3 BC patients shared no somatic mutations with each other and therefore were clonally independent. In other 13 normal lobules, shared mutations including a PIK3CA mutation were observed across multiple adjacent samples, although they were still confined to <10 mm regions. By contrast, we found a large expansion of proliferative lesions sharing a substantial number of somatic mutations with cancer in 5 out of the 6 remaining BC cases. They were expanded over a distance of 25-85 mm, sharing tens to hundreds of mutations including those in BC-related driver genes, such as PIK3CA, AKT1, GATA3, CBFB and PTEN. Of interest, Tumors in 4 out of these 5 cases was Luminal-A type and characterized in common by the presence of 1q gain and 16q loss (1q+/16q-) in both cancer and proliferative lesions. Phylogenetic analysis adapted with the mutation rate in normal cells predicted that 1q+/16q- had been acquired during adolescence to mid-20s, where the clonal expansion had occurred years to decades before the onset of cancer. [Conclusions] Our results suggest that in some BC cases, particularly in those with 1q+/16q-, the clones with the founder driver mutations expanded macroscopically long before the onset of cancer, in which further clonal evolutions recursively occur multi-focally, giving rise to multiple proliferative lesions and ultimately, invasive cancers. Citation Format: Tomomi Nishimura, Nobuyuki Kakiuchi, Kenichi Yoshida, Yasuhide Takeuchi, Hirona Maeda, Yusuke Shiozawa, Masahiro M. Nakagawa, Ryunosuke Saiki, Yotaro Ochi, Tomonori Hirano, Yukiko Kawata, Kosuke Aoki, Masahiro Hirata, Tatsuki R. Kataoka, Takaki Sakurai, Satoko Baba, Yuichi Shiraishi, Kenichi Chiba, Kengo Takeuchi, Hironori Haga, Satoru Miyano, Masakazu Toi, Seishi Ogawa. Clonal evolution of mammary epithelial cells into breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6085.

[Introduction] Proliferative lesions in the breast have been implicated in the development of inv... more [Introduction] Proliferative lesions in the breast have been implicated in the development of invasive breast cancer. Previous studies showed that atypical proliferative lesions and adjacent breast cancers shared common genetic alterations, suggesting that these evolved from the same ancestral cell. However, the clonal structure of atypical proliferative lesions and their clonal dynamics during progression to cancer are poorly understood. In this study, we compared genetic profiles of proliferative lesions and cancer tissues from the same patients to illustrate the clonal evolution of cancer from a non-malignant epithelial cell. [Methods] Multiple samples were collected from various lesions with different disease-level located in the cancer-borne breast, including invasive cancers, using micro-dissection from formalin-fixed, paraffin-embedded surgical specimens. Somatic mutations and copy number alterations (CNAs) were then evaluated by whole exome sequencing. [Results] We analyzed a total of 39 samples from 6 premenopausal females carrying estrogen receptor-positive cancers, where the samples were obtained from non-atypical (N = 1) and atypical (N = 8) proliferative lesions, and non-invasive (N = 21) and invasive (N = 4) cancers. The number of somatic mutations per sample ranged from 2 to 311 and increased with disease progression. Two cases with bilateral cancers had a pathogenic germline mutation of either BRCA2 or TP53, where no somatic mutations or CNAs were shared by individual proliferative lesions, suggesting multifocal independent cancerous evolutions. By contrast, in the remaining four unilateral cases, no pathogenic germline mutations were detected, but all proliferative lesions and cancers, which were separated by a distance of 7- 33 mm, shared one or more driver alterations, such as an AKT1 mutation (UID: KU01), a GATA3 mutation (UID: KU03 and KU06), a CBFB mutation (UID: KU06) and concurrent 1q gain and 16q loss (UID: KU02, KU03 and KU06), while harboring private mutations/CNAs of their own. The phylogenetic analysis based on the number of shared mutations predicted an early origin of these founder genetic alterations, which frequently predated decades before the onset of cancer. [Conclusions] Our results suggest that early breast cancer development is shaped by the evolution of multiple precancerous clones. In hereditary cases, this process is thought to be substantially promoted multi-focally within the entire breasts, due to innate genomic instability in each mammary epithelial cell for pathogenic germline mutations. By contrast, in sporadic cases, the ancestral cell which has acquired a founder genetic alteration expands macroscopically long before the onset of cancer, most likely in early adolescent. These expanded clones work as the niche predisposed to additional mutations, in which branching evolution occurs multi-focally and raises up multiple proliferative lesions, from which invasive cancer finally evolves. Our findings provide unique insight into the early development of breast cancer. Citation Format: Tomomi Nishimura, Kenichi Yoshida, Nobuyuki Kakiuchi, Yasuhide Takeuchi, Yukiko Kawata, Hirona Maeda, Yusuke Shiozawa, Kosuke Aoki, Masahiro Hirata, Tatsuki R Kataoka, Takaki Sakurai, Satoko Baba, Yuichi Shiraishi, Kenichi Chiba, Kengo Takeuchi, Hironori Haga, Satoru Miyano, Masakazu Toi, Seishi Ogawa. Clonal evolution of proliferative lesions into breast cancers [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P4-05-15.

Cancer Research

[Introduction] Proliferative lesions in the breast have been implicated in the development of bre... more [Introduction] Proliferative lesions in the breast have been implicated in the development of breast cancer. Previous studies showed that some proliferative lesions and adjacent breast cancers shared common genetic alterations, suggesting that these originated from the same ancestral cell. However, the clonal structure of normal epithelia and their clonal history during evolution to cancer are poorly understood. In this study, we analyzed genetic profiles of normal epithelia and proliferative lesions in the cancer-borne breast to illustrate the clonal evolution of cancer from a normal epithelial cell. [Methods] Single cell-derived organoids (n=47) were established from breast milk of 4 healthy women aged 22–36 and normal breast tissue of 15 breast cancer patients aged 29–83 to evaluate somatic mutation rate in normal epithelial cells. Multiple normal lobules and proliferative lesions together with cancer lesions were collected using laser-capture micro-dissection (LCM) from fresh frozen (n=3) or formalin-fixed paraffin-embedded (n=5) surgical specimens in 9 premenopausal breast cancer patients. Somatic mutations and copy number alterations were evaluated using whole-genome sequencing. [Results] The mutation profile of single cell-derived organoids suggests that somatic mutations accumulate in normal mammary epithelial cells at a constant rate of 19.4/genome/year before menopause, and the mutation rate decreases to 6.9/genome/year after menopause. Parity was negatively associated with mutation number (-49.3 per life birth). In total, we analyzed 143 LCM samples, including those from 72 normal lobules, 43 proliferative lesions, and 19 non-invasive and 9 invasive cancer samples. Five cases showed a large expansion of proliferative lesions sharing a substantial number of somatic mutations with cancer. These lesions expanded over a distance of 35-90 mm, sharing tens to hundreds of mutations including those in breast cancer-related driver genes, such as PIK3CA, AKT1, GATA3, CBFB and PTEN, while harboring private mutations or copy number alterations of their own. Of interest, the cancers in 4 out of these 5 cases was luminal-A type invasive ducal carcinoma or ER-positive HER2-negative ductal carcinoma in situ, and characterized in common by the presence of der(1;16), concurrent whole-arm 1q gain and 16q loss, in both cancer and proliferative lesions. Phylogenetic analysis adapted with the mutation rate in normal cells predicted that der(1;16) had been acquired between puberty and early 20’s, and the common ancestors of non-cancerous and cancerous lesions emerged by early 30’s, >10 years earlier than at the time of cancer diagnosis. By contrast, analysis of non-cancerous lobules unrelated to cancer showed that der(1;16)-negative non-cancer clones that had emerged after puberty stayed within a single lobule or spatially confined to adjacent lobules and rarely expanded to a large area as observed for those carrying der(1;16), even if the clones had acquired mutations in driver genes such as PIK3CA and PIK3R1, which highlighted the role of der(1;16) in wide clonal expansion. [Conclusions] Our results suggest that in some breast cancer cases, particularly in those with der(1;16), a highly recurrent translocation accounting for the major subset of Luminal A breast cancer, the clones with the funder driver alterations expanded macroscopically long before the onset of cancer, in which further clonal evolutions recursively occur multi-focally, giving rise to multiple proliferative lesions and ultimately, invasive cancers. Our findings provide new insight into the early development of breast cancer. Citation Format: Tomomi Nishimura, Nobuyuki Kakiuchi, Kenichi Yoshida, Takaki Sakurai, Tatsuki R. Kataoka, Eiji Kondoh, Yoshitsugu Chigusa, Masahiko Kawai, Morio Sawada, Takuya Inoue, Yasuhide Takeuchi, Hirona Maeda, Satoko Baba, Yusuke Shiozawa, Ryunosuke Saiki, Masahiro M. Nakagawa, Yasuhito Nannya, Yotaro Ochi, Tomonori Hirano, Yukiko Inagaki-Kawata, Kosuke Aoki, Masahiro Hirata, Eiji Suzuki, Masahiro Takada, Masahiro Kawashima, Kosuke Kawaguchi, Kenichi Chiba, Yuichi Shiraishi, Junko Takita, Satoru Miyano, Masaki Mandai, Kengo Takeuchi, Hironori Haga, Masakazu Toi, Seishi Ogawa. Clonal evolution of mammary epithelial cells into breast cancers [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-13-04.

BMC Medicine

Background Triple-negative breast cancer (TNBC) is a biologically diverse disease, with character... more Background Triple-negative breast cancer (TNBC) is a biologically diverse disease, with characteristics such as homologous recombination deficiency (HRD), gene mutation, and immune reactions. Japan Breast Cancer Research Group 22 is a multicenter trial examining TNBC’s response to neoadjuvant chemotherapy (NAC) according to the HRD status. This translational research investigated the clinical significance of the immune microenvironment of TNBC in association with HRD, tumor BRCA1/2 (tBRCA1/2) mutation, and response to NAC. Methods Patients aged below 65 years with high HRD or germline BRCA1/2 (gBRCA1/2) mutation randomly received paclitaxel + carboplatin (group A1) or eribulin + carboplatin (A2), followed by anthracycline. Patients aged below 65 years with low HRD or those aged 65 years or older without gBRCA1/2 mutation randomly received eribulin + cyclophosphamide (B1) or eribulin + capecitabine (B2); nonresponders to the first four cycles of the therapy received anthracycline. A ...

Breast Cancer Research and Treatment, 2021

Purpose To investigate clinical usefulness of eribulin-based neoadjuvant chemotherapy in triple-n... more Purpose To investigate clinical usefulness of eribulin-based neoadjuvant chemotherapy in triple-negative breast cancer (TNBC) patients. Methods Patients in group A (aged < 65 years with homologous recombination deficiency, HRD, score ≥ 42, or those at any age with germline BRCA mutation, gBRCAm) were randomized to 4 cycles of paclitaxel plus carboplatin (group A1) or eribulin plus carboplatin (group A2), followed by 4 cycles of anthracycline. Patients in group B (aged < 65 years with HRD score < 42, or aged ≥ 65 years without gBRCAm) were randomized to 6 cycles of eribulin plus cyclophosphamide (group B1) or eribulin plus capecitabine (group B2); non-responders to the first 4 cycles of the eribulin-based therapy received anthracycline. Primary endpoint was pCR rate (ypT0-is, ypN0; centrally confirmed). Main secondary endpoint was safety. Results The full analysis set comprised 99 patients. The pCR rate was 65% (90% CI, 46%–81%) and 45% (27%–65%) in groups A1 and A2, respect...

Frontiers in Immunology, 2022

Decidualization of endometrial stromal cells and the presence of immunocompetent cells, including... more Decidualization of endometrial stromal cells and the presence of immunocompetent cells, including human mast cells, play important roles in the establishment of pregnancy. In the present study, the effects of decidualization of endometrial stromal cells on the function of decidual mast cells were elucidated. The in vitro assay revealed that decidualization of an endometrial stromal cell line, T HESCs, increased stem cell factor (SCF) mRNA expression. Decidualization of T HESCs enhanced the production of leukemia inhibitory factor (LIF), and the migration of LAD2 cells when co-cultured with T HESCs and LAD2 cells. In addition, decidualization of T HESCs enhanced cell migration in a human trophoblast cell line, HTR-8/SVneo, increased CD9 expression, a marker for extravillous trophoblast (EVT) differentiation, and decreased the secretion of β human chorionic gonadotropin (hCG), a marker for syncytiotrophoblast (ST) differentiation, when co-cultured with T HESCs, LAD2 cells, and HTR-8/S...

Nature, 2019

Chronic inflammation is accompanied by recurring cycles of tissue destruction and repair and expl... more Chronic inflammation is accompanied by recurring cycles of tissue destruction and repair and explains a major cancer risk 1-3. However, it is poorly understood how such cycles affect clonal composition of tissues, particularly in terms of cancer development. Here we show that in patients with ulcerative colitis (UC), the inflamed intestine undergoes widespread remodelling by pervasive clones positively selected by acquiring mutations commonly involving NFKBIZ, TRAF3IP2, ZC3H12A, PIGR, and HNRNPF, many of which are implicated in downregulation of IL-17 and other proinflammatory signalling. Substantially different mutation profiles between UC-epithelia and cancer indicate distinct mechanisms of positive selection between both. Particularly, NFKBIZ mutations highly prevalent in UC epithelia were rarely found in both sporadic and colitis-associated cancer, suggesting negative selection of NFKBIZ-mutated cells during colorectal carcinogenesis, which was further supported by significantly attenuated colitisinduced tumour formation in Nfkbiz-deficient mice and compromised cell-competition of NFKBIZ-disrupted colorectal cancer cells. Our results highlight common and discrete mechanisms of clonal selection in inflammatory tissues, which unexpectedly reveal cancer vulnerability potentially utilized for therapeutics of colorectal cancer. A Self-archived copy in Kyoto University Research Information Repository https://repository.kulib.kyoto-u.ac.jp A Self-archived copy in Kyoto University Research Information Repository https://repository.kulib.kyoto-u.ac.jp 6 / 47 UC samples weakly but significantly correlated with the interval between diagnosis and sampling (EDFig. 5b,c). Samples from the left-side colon tended to show larger mutation numbers and MCFs, compared with those from the right-side colon (EDFig. 5d,e). To decipher the mechanism of positive selection in UC epithelia, we evaluated dN/dS for 30,062 mutations detected in 399 UC-derived non-dysplastic samples and identified 14 driver genes with evidence of positive selection (dN/dS>1.0, q<0.05) (Fig. 2a,b and EDFig. 6) (Methods), while no positively selected mutations were detected in 179 non-UC samples. Mutations in most of these drivers in UC epithelium were predicted to cause a protein truncation and loss-offunction. Exceptions were mutations in ZC3H12A, KRAS, and HNRNPF, which exhibited prominent mutational hotspots indicative of some neomorphic functions (Fig. 2a and EDFig. 6a). NFKBIZ, TRAF3IP2, and RNF43 frequently underwent biallelic inactivation, which was less common in other drivers (Fig. 2b). A single bulk-crypt sample may contain multiple independent clones and subclones carrying discrete driver mutations. Thus, to precisely determine the frequency of mutations in UC epithelia on the basis of independent clones, we first clustered mutations detected in bulk-crypt samples using PyClone and based on the cell fraction predicted for each mutation cluster, identified a total of 183 independent clones according to the 'Pigeonhole principle' (EDFig. 1d) (Methods). Among these, 166 (90%) harboured one or more driver mutations, which most frequently affected NFKBIZ, followed by ARID1A, PIGR, KRAS, and ZC3H12A (Fig. 2b). Conspicuously, the molecules mutated in UC epithelia displayed a surprising overlap to signalling pathways converging into NFKBIZ (also known as IκBζ) and related molecules (ZC3H12A and PIGR) (Fig. 2c), where the mutations in these molecules occurred in a mutually exclusive manner (Fig. 2b). Among these mutational targets most prominent was the molecules involved in IL-17 signalling, including IL-17 receptors (IL17RA/IL17RC) and its adaptor, TRAF3IP2, which were shown to be positively selected in UC epithelia (Methods), suggesting a major role of IL-17 signalling (Fig.2c). Moreover, other inflammation-related cytokine receptors and pattern recognition receptors (PRRs) implicated in NFKBIZ upregulation, including OSMR, LTBR, IL20RA, and TLR9 12 (EDFig. 6b), also underwent loss-of-function mutations in occasional clones (Methods). Expansion of clones carrying these cytokine/PRR pathway mutations was intensively evaluated in surgically obtained rectum specimens from 6 UC patients using densely collected samples (Fig. 2d and EDFig. 7a). We identified 41 independent driver-mutated clones that involved ≥2 samples in these specimens, of which 31 had the mutations in the above pathways. Conspicuously, in these patients, 53-83% of the entire rectum was replaced by clones having one or more driver mutations, most of which were explained by cytokine/PRR pathway mutations (Fig. 2e). The expansion could be extensive, occupying an epithelial area as large as 19 cm 2 in A Self-archived copy in