Ku70, a DNA repair protein, binds to the damaged DNA ends and orchestrates the recruitment of other proteins to facilitate repair of DNA double-strand breaks. Besides its essential role in DNA repair, several studies have highlighted non-classical functions of Ku70 in cellular processes. However, its function in immune homeostasis and anti-tumor immunity remains unknown. Here, we discovered a marked association between elevated Ku70 expression and unfavorable prognosis in lung adenocarcinoma, focusing specifically on increased Ku70 levels in tumor-infiltrated Treg cells. Using a lung-colonizing tumor model of in mice with Treg-specific Ku70 deficiency, we demonstrated that deletion of Ku70 in Treg cells led to a stronger anti-tumor response and slower tumor growth due to impaired immune-suppressive capacity of Treg cells. Furthermore, we confirmed that Ku70 played a critical role in sustaining the suppressive function of human Treg cells. We found that Ku70 bound to FOXP3 and occupied FOXP3-bound genomic sites to support its transcriptional activities. These findings not only unveil a non-homologous end joining (NHEJ)-independent role of Ku70 crucial for Treg suppressive function, but also underscore the potential of targeting Ku70 as an effective strategy in cancer therapy, aiming to both restrain cancer cells and enhance pulmonary anti-tumor immunity.
{"title":"Non-classical action of Ku70 promotes Treg suppressive function through a FOXP3-dependent mechanism in lung adenocarcinoma.","authors":"Qianru Huang,Na Tian,Jianfeng Zhang,Shiyang Song,Hao Cheng,Xinnan Liu,Wenle Zhang,Youqiong Ye,Yanhua Du,Xueyu Dai,Rui Liang,Dan Li,Sheng-Ming Dai,Chuan Wang,Zhi Chen,Qianjun Zhou,Bin Li","doi":"10.1172/jci178079","DOIUrl":"https://doi.org/10.1172/jci178079","url":null,"abstract":"Ku70, a DNA repair protein, binds to the damaged DNA ends and orchestrates the recruitment of other proteins to facilitate repair of DNA double-strand breaks. Besides its essential role in DNA repair, several studies have highlighted non-classical functions of Ku70 in cellular processes. However, its function in immune homeostasis and anti-tumor immunity remains unknown. Here, we discovered a marked association between elevated Ku70 expression and unfavorable prognosis in lung adenocarcinoma, focusing specifically on increased Ku70 levels in tumor-infiltrated Treg cells. Using a lung-colonizing tumor model of in mice with Treg-specific Ku70 deficiency, we demonstrated that deletion of Ku70 in Treg cells led to a stronger anti-tumor response and slower tumor growth due to impaired immune-suppressive capacity of Treg cells. Furthermore, we confirmed that Ku70 played a critical role in sustaining the suppressive function of human Treg cells. We found that Ku70 bound to FOXP3 and occupied FOXP3-bound genomic sites to support its transcriptional activities. These findings not only unveil a non-homologous end joining (NHEJ)-independent role of Ku70 crucial for Treg suppressive function, but also underscore the potential of targeting Ku70 as an effective strategy in cancer therapy, aiming to both restrain cancer cells and enhance pulmonary anti-tumor immunity.","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"168 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142490552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yurong Song,Ryan N Baugher,Todd B Young,Brandon Somerville,Yuriko Mori,Ligia A Pinto,Kim E Nichols,Robert H Shoemaker
{"title":"Frameshift mutation spectra overlap between constitutional mismatch repair deficiency tumors and Lynch syndrome tumors.","authors":"Yurong Song,Ryan N Baugher,Todd B Young,Brandon Somerville,Yuriko Mori,Ligia A Pinto,Kim E Nichols,Robert H Shoemaker","doi":"10.1172/jci185999","DOIUrl":"https://doi.org/10.1172/jci185999","url":null,"abstract":"","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"234 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488295","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jiawei Wang,Huogang Wang,Wei Zhou,Xin Luo,Huijuan Wang,Qing Meng,Jiaxin Chen,Xiaoyu Chen,Yinqiang Liu,David W Chan,Zhenyu Ju,Zhangfa Song
BRAFV600E-mutant metastatic colorectal cancer (mCRC) is associated with poor prognosis. The combination of anti-BRAF/EGFR (encorafenib/cetuximab) treatment for patients with BRAFV600E-mutant mCRC improved clinical benefits; unfortunately, inevitable acquired resistance limits the treatment outcome, and the mechanism has not been validated. Here, we discovered that monoacylglycerol O-Acyltransferase 3 (MOGAT3) mediated diacylglycerol (DAG) accumulation contributed to acquired resistance to encorafenib/cetuximab by dissecting BRAFV600E-mutant mCRC patient-derived xenograft (PDX) model exposed to encorafenib/cetuximab administration. Mechanistically, upregulated MOGAT3 promotes DAG synthesis and reduces fatty acid oxidation (FAO)-promoting DAG accumulation and activating PKCα-CRAF-MEK-ERK, driving acquired resistance. Resistance-induced hypoxia promotes MOGAT3 transcriptional elevation; simultaneously, MOGAT3-mediated DAG accumulation increases HIF1A expression in translation level through PKCα-CRAF-eIF4E activation, strengthening the resistance status. Intriguingly, reducing intratumoral DAG by fenofibrate or Pf-06471553 restores the antitumor efficacy of encorafenib/cetuximab on resistant BRAFV600E-mutant mCRC, interrupted PKCα-CRAF-MEK-ERK signaling. These findings reveal the critical metabolite DAG as a modulator of encorafenib/cetuximab efficacy in BRAFV600E-mutant mCRC, suggesting that fenofibrate may prove beneficial for resistant BRAFV600E-mutant mCRC patients.
BRAFV600E突变的转移性结直肠癌（mCRC）预后较差。对BRAFV600E突变的mCRC患者进行抗BRAF/EGFR（安戈非尼/西妥昔单抗）联合治疗可提高临床疗效；但遗憾的是，不可避免的获得性耐药限制了治疗效果，其机制尚未得到验证。在这里，我们通过剖析暴露于安戈非尼/西妥昔单抗治疗的BRAFV600E突变型mCRC患者异种移植（PDX）模型，发现单酰基甘油O-酰基转移酶3（MOGAT3）介导的二酰基甘油（DAG）积累导致了对安戈非尼/西妥昔单抗的获得性耐药。从机理上讲，上调的MOGAT3促进了DAG的合成并减少了脂肪酸氧化（FAO）--促进了DAG的积累并激活了PKCα-CRAF-MEK-ERK，从而推动了获得性耐药性的产生。耐药性诱导的低氧促进了 MOGAT3 的转录升高；同时，MOGAT3 介导的 DAG 积累通过 PKCα-CRAF-eIF4E 激活，在翻译水平上增加了 HIF1A 的表达，从而加强了耐药性状态。有趣的是，通过非诺贝特或 Pf-06471553 减少瘤内 DAG 可恢复安戈非尼/西妥昔单抗对耐药 BRAFV600E 突变 mCRC 的抗肿瘤疗效，从而中断 PKCα-CRAF-MEK-ERK 信号传导。这些发现揭示了关键代谢物DAG是安戈非尼/西妥昔单抗对BRAFV600E突变型mCRC疗效的调节剂，表明非诺贝特可能对耐药的BRAFV600E突变型mCRC患者有益。
{"title":"MOGAT3-Mediated DAG Accumulation Drives Acquired Resistance to Anti-BRAF/EGFR Therapy in BRAFV600E-Mutant Metastatic Colorectal Cancer.","authors":"Jiawei Wang,Huogang Wang,Wei Zhou,Xin Luo,Huijuan Wang,Qing Meng,Jiaxin Chen,Xiaoyu Chen,Yinqiang Liu,David W Chan,Zhenyu Ju,Zhangfa Song","doi":"10.1172/jci182217","DOIUrl":"https://doi.org/10.1172/jci182217","url":null,"abstract":"BRAFV600E-mutant metastatic colorectal cancer (mCRC) is associated with poor prognosis. The combination of anti-BRAF/EGFR (encorafenib/cetuximab) treatment for patients with BRAFV600E-mutant mCRC improved clinical benefits; unfortunately, inevitable acquired resistance limits the treatment outcome, and the mechanism has not been validated. Here, we discovered that monoacylglycerol O-Acyltransferase 3 (MOGAT3) mediated diacylglycerol (DAG) accumulation contributed to acquired resistance to encorafenib/cetuximab by dissecting BRAFV600E-mutant mCRC patient-derived xenograft (PDX) model exposed to encorafenib/cetuximab administration. Mechanistically, upregulated MOGAT3 promotes DAG synthesis and reduces fatty acid oxidation (FAO)-promoting DAG accumulation and activating PKCα-CRAF-MEK-ERK, driving acquired resistance. Resistance-induced hypoxia promotes MOGAT3 transcriptional elevation; simultaneously, MOGAT3-mediated DAG accumulation increases HIF1A expression in translation level through PKCα-CRAF-eIF4E activation, strengthening the resistance status. Intriguingly, reducing intratumoral DAG by fenofibrate or Pf-06471553 restores the antitumor efficacy of encorafenib/cetuximab on resistant BRAFV600E-mutant mCRC, interrupted PKCα-CRAF-MEK-ERK signaling. These findings reveal the critical metabolite DAG as a modulator of encorafenib/cetuximab efficacy in BRAFV600E-mutant mCRC, suggesting that fenofibrate may prove beneficial for resistant BRAFV600E-mutant mCRC patients.","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"63 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ru M Wen,Jessica C Stark,G Edward W Marti,Zenghua Fan,Aram Lyu,Fernando Jose Garcia Marques,Xiangyue Zhang,Nicholas M Riley,Sarah M Totten,Abel Bermudez,Rosalie Nolley,Hongjuan Zhao,Lawrence Fong,Edgar G Engleman,Sharon J Pitteri,Carolyn R Bertozzi,James D Brooks
Prostate cancer is the second leading cause of male cancer death in the U.S. Current immune checkpoint inhibitor-based immunotherapies have improved survival for many malignancies; however, they have failed to prolong survival for prostate cancer. Siglecs (sialic acid-binding immunoglobulin-like lectins) are expressed on immune cells and regulate immune responses and function. Siglec-7 and Siglec-9 contribute to immune evasion by interacting with their ligands. However, the role of Siglec-7/9 receptors and their ligands in prostate cancer remains poorly understood. Here, we find that Siglec-7 and Siglec-9 are associated with poor prognosis in prostate cancer patients, and are highly expressed in myeloid cells, including macrophages, in prostate tumor tissues. Siglecs-7 and -9 ligands were expressed in prostate cancer cells and human prostate tumor tissues. Blocking the interactions between Siglec-7/9 and sialic acids inhibited prostate cancer xenograft growth and increased immune cell infiltration in humanized mice in vivo. Using a CRISPRi screen and mass spectrometry, we identified CD59 as a candidate Siglec-9 ligand in prostate cancer. The identification of Siglecs-7 and -9 as potential therapeutic targets, including CD59/Siglec-9 axis, opens up opportunities for immune-based interventions in prostate cancer.
{"title":"Sialylated glycoproteins suppress immune cell killing by binding to Siglec-7 and Siglec-9 in prostate cancer.","authors":"Ru M Wen,Jessica C Stark,G Edward W Marti,Zenghua Fan,Aram Lyu,Fernando Jose Garcia Marques,Xiangyue Zhang,Nicholas M Riley,Sarah M Totten,Abel Bermudez,Rosalie Nolley,Hongjuan Zhao,Lawrence Fong,Edgar G Engleman,Sharon J Pitteri,Carolyn R Bertozzi,James D Brooks","doi":"10.1172/jci180282","DOIUrl":"https://doi.org/10.1172/jci180282","url":null,"abstract":"Prostate cancer is the second leading cause of male cancer death in the U.S. Current immune checkpoint inhibitor-based immunotherapies have improved survival for many malignancies; however, they have failed to prolong survival for prostate cancer. Siglecs (sialic acid-binding immunoglobulin-like lectins) are expressed on immune cells and regulate immune responses and function. Siglec-7 and Siglec-9 contribute to immune evasion by interacting with their ligands. However, the role of Siglec-7/9 receptors and their ligands in prostate cancer remains poorly understood. Here, we find that Siglec-7 and Siglec-9 are associated with poor prognosis in prostate cancer patients, and are highly expressed in myeloid cells, including macrophages, in prostate tumor tissues. Siglecs-7 and -9 ligands were expressed in prostate cancer cells and human prostate tumor tissues. Blocking the interactions between Siglec-7/9 and sialic acids inhibited prostate cancer xenograft growth and increased immune cell infiltration in humanized mice in vivo. Using a CRISPRi screen and mass spectrometry, we identified CD59 as a candidate Siglec-9 ligand in prostate cancer. The identification of Siglecs-7 and -9 as potential therapeutic targets, including CD59/Siglec-9 axis, opens up opportunities for immune-based interventions in prostate cancer.","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"51 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Laura Ondrisova,Vaclav Seda,Krystof Hlavac,Petra Pavelkova,Eva Hoferkova,Giorgia Chiodin,Lenka Kostalova,Gabriela Mladonicka Pavlasova,Daniel Filip,Josef Vecera,Pedro Faria Zeni,Jan Oppelt,Zuzana Kahounova,Rachel Vichova,Karel Soucek,Anna Panovska,Karla Plevova,Sarka Pospisilova,Martin Simkovic,Filip Vrbacky,Daniel Lysak,Stacey M Fernandes,Matthew S Davids,Alba Maiques-Diaz,Stella Charalampopoulou,Jose I Martin-Subero,Jennifer R Brown,Michael Doubek,Francesco Forconi,Jiri Mayer,Marek Mraz
BTK inhibitor therapy induces peripheral blood lymphocytosis in chronic lymphocytic leukemia (CLL) lasting for several months. It remains unclear whether non-genetic adaptation mechanisms exist, allowing CLL cells' survival during BTK inhibitor-induced lymphocytosis and/or playing a role in therapy resistance. We show that in approximately 70 % of CLL cases, ibrutinib treatment in vivo increases Akt activity above pre-therapy levels within several weeks, leading to compensatory CLL cell survival and a more prominent lymphocytosis on therapy. Ibrutinib-induced Akt phosphorylation (pAktS473) is caused by the upregulation of FoxO1 transcription factor, which induces expression of Rictor, an assembly protein for mTORC2 protein complex that directly phosphorylates Akt at serine 473 (S473). Knock-out or inhibition of FoxO1 or Rictor led to a dramatic decrease in Akt phosphorylation and growth disadvantage for malignant B cells in the presence of ibrutinib (or PI3K inhibitor idelalisib) in vitro and in vivo. FoxO1/Rictor/pAktS473 axis represents an early non-genetic adaptation to BCR inhibitor therapy not requiring PI3Kδ or BTK kinase activity. We further demonstrate that FoxO1 can be targeted therapeutically, and its inhibition induces CLL cells' apoptosis alone or in combination with BTK inhibitors (ibrutinib, acalabrutinib, pirtobrutinib) and blocks their proliferation triggered by T-cell factors (CD40L, IL-4, and IL-21).
{"title":"FoxO1/Rictor axis induces a non-genetic adaptation to Ibrutinib via Akt activation in chronic lymphocytic leukemia.","authors":"Laura Ondrisova,Vaclav Seda,Krystof Hlavac,Petra Pavelkova,Eva Hoferkova,Giorgia Chiodin,Lenka Kostalova,Gabriela Mladonicka Pavlasova,Daniel Filip,Josef Vecera,Pedro Faria Zeni,Jan Oppelt,Zuzana Kahounova,Rachel Vichova,Karel Soucek,Anna Panovska,Karla Plevova,Sarka Pospisilova,Martin Simkovic,Filip Vrbacky,Daniel Lysak,Stacey M Fernandes,Matthew S Davids,Alba Maiques-Diaz,Stella Charalampopoulou,Jose I Martin-Subero,Jennifer R Brown,Michael Doubek,Francesco Forconi,Jiri Mayer,Marek Mraz","doi":"10.1172/jci173770","DOIUrl":"https://doi.org/10.1172/jci173770","url":null,"abstract":"BTK inhibitor therapy induces peripheral blood lymphocytosis in chronic lymphocytic leukemia (CLL) lasting for several months. It remains unclear whether non-genetic adaptation mechanisms exist, allowing CLL cells' survival during BTK inhibitor-induced lymphocytosis and/or playing a role in therapy resistance. We show that in approximately 70 % of CLL cases, ibrutinib treatment in vivo increases Akt activity above pre-therapy levels within several weeks, leading to compensatory CLL cell survival and a more prominent lymphocytosis on therapy. Ibrutinib-induced Akt phosphorylation (pAktS473) is caused by the upregulation of FoxO1 transcription factor, which induces expression of Rictor, an assembly protein for mTORC2 protein complex that directly phosphorylates Akt at serine 473 (S473). Knock-out or inhibition of FoxO1 or Rictor led to a dramatic decrease in Akt phosphorylation and growth disadvantage for malignant B cells in the presence of ibrutinib (or PI3K inhibitor idelalisib) in vitro and in vivo. FoxO1/Rictor/pAktS473 axis represents an early non-genetic adaptation to BCR inhibitor therapy not requiring PI3Kδ or BTK kinase activity. We further demonstrate that FoxO1 can be targeted therapeutically, and its inhibition induces CLL cells' apoptosis alone or in combination with BTK inhibitors (ibrutinib, acalabrutinib, pirtobrutinib) and blocks their proliferation triggered by T-cell factors (CD40L, IL-4, and IL-21).","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"17 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Allison Joy Nielsen,Gabriella Kyra Albert,Amelia Sanchez,Jiangli Chen,Jing Liu,Andres Sebastian Davalos,Degui Geng,Xander G Bradeen,Jennifer D Hintzsche,William Robinson,Martin McCarter,Carol M Amato,Richard Tobin,Kasey L Couts,Breelyn Ann Wilky,Eduardo Davila
Effective antitumor T cell activity relies on the expression and MHC presentation of tumor neoantigens. Tumor cells can evade T cell detection by silencing the transcription of antigens or by altering MHC machinery resulting in inadequate neoantigen-specific T cell activation. We identified DNA-PK inhibitor (DNA-PKi) NU7441 as a promising immunomodulator that reduced immunosuppressive proteins while increasing MHC-I expression in a panel of human melanoma cell lines. In tumor-bearing mice, combination therapy using NU7441 and immune adjuvants STING ligand and CD40 agonist (NU-SL40) substantially increased and diversified the neoantigen landscape, antigen presenting machinery, and consequently substantially increased both the number and repertoire of neoantigen-reactive tumor infiltrating lymphocytes (TILs). DNA-PK-inhibition or knockout promoted transcription and protein expression of various neoantigens in human and mouse melanomas and induced sensitivity to ICB in resistant tumors. In patients, PRKDC levels inversely correlated with MHC I expression and CD8 TILs but positively correlated with increased neoantigen loads and improved responses to ICB. These studies suggest that inhibiting DNA-PK activity can restore tumor immunogenicity by increasing neoantigen expression and presentation and broadening the neoantigen-reactive T cell population.
有效的抗肿瘤 T 细胞活性依赖于肿瘤新抗原的表达和 MHC 呈递。肿瘤细胞可以通过抑制抗原转录或改变 MHC 机制来逃避 T 细胞检测，从而导致新抗原特异性 T 细胞活化不足。我们发现DNA-PK抑制剂（DNA-PKi）NU7441是一种很有前景的免疫调节剂，它能减少免疫抑制蛋白，同时增加人类黑色素瘤细胞系的MHC-I表达。在肿瘤小鼠中，使用 NU7441 和免疫佐剂 STING 配体及 CD40 激动剂（NU-SL40）进行联合治疗，可显著增加新抗原的分布和抗原递呈机制的多样性，从而大幅增加新抗原反应性肿瘤浸润淋巴细胞（TILs）的数量和种类。DNA-PK 抑制或基因敲除可促进人和小鼠黑色素瘤中各种新抗原的转录和蛋白表达，并诱导耐药肿瘤对 ICB 敏感。在患者中，PRKDC水平与MHC I表达和CD8 TILs成反比，但与新抗原负荷的增加和对ICB反应的改善成正比。这些研究表明，抑制 DNA-PK 活性可以通过增加新抗原表达和呈现以及扩大新抗原反应性 T 细胞群来恢复肿瘤免疫原性。
{"title":"DNA-PK inhibition enhances neoantigen diversity and increases T cell responses to immunoresistant tumors.","authors":"Allison Joy Nielsen,Gabriella Kyra Albert,Amelia Sanchez,Jiangli Chen,Jing Liu,Andres Sebastian Davalos,Degui Geng,Xander G Bradeen,Jennifer D Hintzsche,William Robinson,Martin McCarter,Carol M Amato,Richard Tobin,Kasey L Couts,Breelyn Ann Wilky,Eduardo Davila","doi":"10.1172/jci180278","DOIUrl":"https://doi.org/10.1172/jci180278","url":null,"abstract":"Effective antitumor T cell activity relies on the expression and MHC presentation of tumor neoantigens. Tumor cells can evade T cell detection by silencing the transcription of antigens or by altering MHC machinery resulting in inadequate neoantigen-specific T cell activation. We identified DNA-PK inhibitor (DNA-PKi) NU7441 as a promising immunomodulator that reduced immunosuppressive proteins while increasing MHC-I expression in a panel of human melanoma cell lines. In tumor-bearing mice, combination therapy using NU7441 and immune adjuvants STING ligand and CD40 agonist (NU-SL40) substantially increased and diversified the neoantigen landscape, antigen presenting machinery, and consequently substantially increased both the number and repertoire of neoantigen-reactive tumor infiltrating lymphocytes (TILs). DNA-PK-inhibition or knockout promoted transcription and protein expression of various neoantigens in human and mouse melanomas and induced sensitivity to ICB in resistant tumors. In patients, PRKDC levels inversely correlated with MHC I expression and CD8 TILs but positively correlated with increased neoantigen loads and improved responses to ICB. These studies suggest that inhibiting DNA-PK activity can restore tumor immunogenicity by increasing neoantigen expression and presentation and broadening the neoantigen-reactive T cell population.","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488302","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ye Ji Choi,Gabriel Richard,Guanshi Zhang,Jeffrey B Hodgin,Dawit S Demeke,Yingbao Yang,Jennifer A Schaub,Ian M Tamayo,Bhupendra K Gurung,Abhijit S Naik,Viji Nair,Carissa Birznieks,Alexis MacDonald,Phoom Narongkiatikhun,Susan Gross,Lynette Driscoll,Maureen Flynn,Kalie Tommerdahl,Kristen J Nadeau,Viral N Shah,Tim Vigers,Janet K Snell-Bergeon,Jessica Kendrick,Daniel H van Raalte,Lu-Ping Li,Pottumarthi Prasad,Patricia Ladd,Bennett B Chin,David Z Cherney,Phillip J McCown,Fadhl Alakwaa,Edgar A Otto,Frank C Brosius,Pierre Jean Saulnier,Victor G Puelles,Jesse A Goodrich,Kelly Street,Manjeri A Venkatachalam,Aaron Ruiz,Ian H de Boer,Robert G Nelson,Laura Pyle,Denis P Blondin,Kumar Sharma,Matthias Kretzler,Petter Bjornstad
BACKGROUNDIn type 1 diabetes (T1D), impaired insulin sensitivity may contribute to the development of diabetic kidney disease (DKD) through alterations in kidney oxidative metabolism.METHODSYoung adults with T1D (n = 30) and healthy controls (HC, n = 20) underwent hyperinsulinemic-euglycemic clamp studies, MRI, 11C-acetate PET, kidney biopsies, single-cell RNA sequencing, and spatial metabolomics to assess this relationship.RESULTSParticipants with T1D had significantly higher glomerular basement membrane thickness compared to HC. T1D participants exhibited lower insulin sensitivity and cortical oxidative metabolism, correlating with higher insulin sensitivity. Proximal tubular transcripts of TCA cycle and oxidative phosphorylation enzymes were lower in T1D. Spatial metabolomics showed reductions in tubular TCA cycle intermediates, indicating mitochondrial dysfunction. The Slingshot algorithm identified a lineage of proximal tubular cells progressing from stable to adaptive/maladaptive subtypes, using pseudotime trajectory analysis, which computationally orders cells along a continuum of states. This analysis revealed distinct distribution patterns between T1D and HC, with attenuated oxidative metabolism in T1D attributed to a greater proportion of adaptive/maladaptive subtypes with low expression of TCA cycle and oxidative phosphorylation transcripts. Pseudotime progression associated with higher HbA1c, BMI, GBM, and lower insulin sensitivity and cortical oxidative metabolism.CONCLUSIONThese early structural and metabolic changes in T1D kidneys may precede clinical DKD.TRIAL REGISTRATIONCLINICALTRIALSgov NCT04074668.
{"title":"Attenuated kidney oxidative metabolism in young adults with type 1 diabetes.","authors":"Ye Ji Choi,Gabriel Richard,Guanshi Zhang,Jeffrey B Hodgin,Dawit S Demeke,Yingbao Yang,Jennifer A Schaub,Ian M Tamayo,Bhupendra K Gurung,Abhijit S Naik,Viji Nair,Carissa Birznieks,Alexis MacDonald,Phoom Narongkiatikhun,Susan Gross,Lynette Driscoll,Maureen Flynn,Kalie Tommerdahl,Kristen J Nadeau,Viral N Shah,Tim Vigers,Janet K Snell-Bergeon,Jessica Kendrick,Daniel H van Raalte,Lu-Ping Li,Pottumarthi Prasad,Patricia Ladd,Bennett B Chin,David Z Cherney,Phillip J McCown,Fadhl Alakwaa,Edgar A Otto,Frank C Brosius,Pierre Jean Saulnier,Victor G Puelles,Jesse A Goodrich,Kelly Street,Manjeri A Venkatachalam,Aaron Ruiz,Ian H de Boer,Robert G Nelson,Laura Pyle,Denis P Blondin,Kumar Sharma,Matthias Kretzler,Petter Bjornstad","doi":"10.1172/jci183984","DOIUrl":"https://doi.org/10.1172/jci183984","url":null,"abstract":"BACKGROUNDIn type 1 diabetes (T1D), impaired insulin sensitivity may contribute to the development of diabetic kidney disease (DKD) through alterations in kidney oxidative metabolism.METHODSYoung adults with T1D (n = 30) and healthy controls (HC, n = 20) underwent hyperinsulinemic-euglycemic clamp studies, MRI, 11C-acetate PET, kidney biopsies, single-cell RNA sequencing, and spatial metabolomics to assess this relationship.RESULTSParticipants with T1D had significantly higher glomerular basement membrane thickness compared to HC. T1D participants exhibited lower insulin sensitivity and cortical oxidative metabolism, correlating with higher insulin sensitivity. Proximal tubular transcripts of TCA cycle and oxidative phosphorylation enzymes were lower in T1D. Spatial metabolomics showed reductions in tubular TCA cycle intermediates, indicating mitochondrial dysfunction. The Slingshot algorithm identified a lineage of proximal tubular cells progressing from stable to adaptive/maladaptive subtypes, using pseudotime trajectory analysis, which computationally orders cells along a continuum of states. This analysis revealed distinct distribution patterns between T1D and HC, with attenuated oxidative metabolism in T1D attributed to a greater proportion of adaptive/maladaptive subtypes with low expression of TCA cycle and oxidative phosphorylation transcripts. Pseudotime progression associated with higher HbA1c, BMI, GBM, and lower insulin sensitivity and cortical oxidative metabolism.CONCLUSIONThese early structural and metabolic changes in T1D kidneys may precede clinical DKD.TRIAL REGISTRATIONCLINICALTRIALSgov NCT04074668.","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"67 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Saiti S Halder,Michael J Rynkiewicz,Lynne Kim,Meaghan Barry,Ahmed Ga Zied,Lorenzo R Sewanan,Jonathan A Kirk,Jeffrey R Moore,William Lehman,Stuart G Campbell
Hypertrophic and dilated cardiomyopathies (HCM and DCM, respectively) are inherited disorders that may be caused by mutations to the same sarcomeric protein but have completely different clinical phenotypes. The precise mechanisms by which point mutations within the same gene bring about phenotypic diversity remain unclear. Our objective has been to develop a mechanistic explanation of diverging phenotypes in two TPM1 mutations, E62Q (HCM) and E54K (DCM). Drawing on data from the literature and experiments with stem cell-derived cardiomyocytes expressing the TPM1 mutations of interest, we constructed computational simulations that provide plausible explanations of the distinct muscle contractility caused by each variant. In E62Q, increased calcium sensitivity and hypercontractility was explained most accurately by a reduction in effective molecular stiffness of tropomyosin and alterations in its interactions with the actin thin filament that favor the 'closed' regulatory state. By contrast, the E54K mutation appeared to act via long-range allosteric interactions to increase the association rate of the C-terminal troponin I mobile domain to tropomyosin/actin. These mutation-linked molecular events produced diverging alterations in gene expression that can be observed in human engineered heart tissues. Modulators of myosin activity confirmed our proposed mechanisms by rescuing normal contractile behavior in accordance with predictions.
肥厚型心肌病和扩张型心肌病（分别为 HCM 和 DCM）是一种遗传性疾病，可能由相同的肌浆蛋白突变引起，但临床表型却完全不同。同一基因中的点突变导致表型多样性的确切机制仍不清楚。我们的目标是对两种 TPM1 突变（E62Q（HCM）和 E54K（DCM））的不同表型做出机理解释。我们利用文献数据和表达相关 TPM1 突变的干细胞衍生心肌细胞的实验，构建了计算模拟，为每种变异引起的不同肌肉收缩能力提供了合理的解释。在 E62Q 突变体中，钙敏感性和过度收缩性增加的最准确解释是肌球蛋白有效分子刚度的降低及其与肌动蛋白细丝相互作用的改变，这种改变有利于 "封闭 "调节状态。相比之下，E54K 突变似乎是通过长程异构相互作用来增加 C 端肌钙蛋白 I 移动结构域与肌球蛋白/肌动蛋白的结合率。这些与突变相关的分子事件导致了基因表达的不同变化，这些变化可以在人体工程心脏组织中观察到。肌球蛋白活性调节剂根据预测挽救了正常的收缩行为，从而证实了我们提出的机制。
{"title":"Distinct mechanisms drive divergent phenotypes in hypertrophic and dilated cardiomyopathy associated TPM1 variants.","authors":"Saiti S Halder,Michael J Rynkiewicz,Lynne Kim,Meaghan Barry,Ahmed Ga Zied,Lorenzo R Sewanan,Jonathan A Kirk,Jeffrey R Moore,William Lehman,Stuart G Campbell","doi":"10.1172/jci179135","DOIUrl":"https://doi.org/10.1172/jci179135","url":null,"abstract":"Hypertrophic and dilated cardiomyopathies (HCM and DCM, respectively) are inherited disorders that may be caused by mutations to the same sarcomeric protein but have completely different clinical phenotypes. The precise mechanisms by which point mutations within the same gene bring about phenotypic diversity remain unclear. Our objective has been to develop a mechanistic explanation of diverging phenotypes in two TPM1 mutations, E62Q (HCM) and E54K (DCM). Drawing on data from the literature and experiments with stem cell-derived cardiomyocytes expressing the TPM1 mutations of interest, we constructed computational simulations that provide plausible explanations of the distinct muscle contractility caused by each variant. In E62Q, increased calcium sensitivity and hypercontractility was explained most accurately by a reduction in effective molecular stiffness of tropomyosin and alterations in its interactions with the actin thin filament that favor the 'closed' regulatory state. By contrast, the E54K mutation appeared to act via long-range allosteric interactions to increase the association rate of the C-terminal troponin I mobile domain to tropomyosin/actin. These mutation-linked molecular events produced diverging alterations in gene expression that can be observed in human engineered heart tissues. Modulators of myosin activity confirmed our proposed mechanisms by rescuing normal contractile behavior in accordance with predictions.","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"234 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ting-Yu Chang,Yan Yan,Zih-Yao Yu,Moeez Rathore,Nian-Zhe Lee,Hui-Ju Tseng,Li-Hsin Cheng,Wei-Jan Huang,Wei Zhang,Ernest R Chan,Yulan Qing,Ming-Lun Kang,Rui Wang,Kelvin K Tsai,John J Pink,William E Harte,Stanton L Gerson,Sung-Bau Lee
The elevated level of replication stress is an intrinsic characteristic of cancer cells. Targeting the mechanisms that maintain genome stability to further increase replication stress and thus induce severe genome instability has become a promising approach for cancer treatment. Here, we identify histone deacetylase 8 (HDAC8) as a drug target whose inactivation synergizes with the inhibition of checkpoint kinases to elicit substantial replication stress and compromise genome integrity selectively in cancer cells. We showed that simultaneous inhibition of HDAC8 and checkpoint kinases led to extensive replication fork collapse, irreversible cell-cycle arrest, and synergistic vulnerability in various cancer cells. The efficacy of the combination treatment was further validated in patient tumor-derived organoid (PDO) and xenograft mouse (PDX) models, providing important insights into patient-specific drug responses. Our data revealed that HDAC8 activity was essential for reducing the acetylation level of structural maintenance of chromosomes protein 3 (SMC3) ahead of replication forks and preventing R loop formation. HDAC8 inactivation resulted in slowed fork progression and checkpoint kinase activation. Our findings indicate that HDAC8 guards the integrity of the replicating genome, and the cancer-specific synthetic lethality between HDAC8 and checkpoint kinases provides a promising replication stress-targeting strategy for treating a broad range of cancers.
{"title":"Combined HDAC8 and checkpoint kinase inhibition induces tumor-selective synthetic lethality in preclinical models.","authors":"Ting-Yu Chang,Yan Yan,Zih-Yao Yu,Moeez Rathore,Nian-Zhe Lee,Hui-Ju Tseng,Li-Hsin Cheng,Wei-Jan Huang,Wei Zhang,Ernest R Chan,Yulan Qing,Ming-Lun Kang,Rui Wang,Kelvin K Tsai,John J Pink,William E Harte,Stanton L Gerson,Sung-Bau Lee","doi":"10.1172/jci165448","DOIUrl":"https://doi.org/10.1172/jci165448","url":null,"abstract":"The elevated level of replication stress is an intrinsic characteristic of cancer cells. Targeting the mechanisms that maintain genome stability to further increase replication stress and thus induce severe genome instability has become a promising approach for cancer treatment. Here, we identify histone deacetylase 8 (HDAC8) as a drug target whose inactivation synergizes with the inhibition of checkpoint kinases to elicit substantial replication stress and compromise genome integrity selectively in cancer cells. We showed that simultaneous inhibition of HDAC8 and checkpoint kinases led to extensive replication fork collapse, irreversible cell-cycle arrest, and synergistic vulnerability in various cancer cells. The efficacy of the combination treatment was further validated in patient tumor-derived organoid (PDO) and xenograft mouse (PDX) models, providing important insights into patient-specific drug responses. Our data revealed that HDAC8 activity was essential for reducing the acetylation level of structural maintenance of chromosomes protein 3 (SMC3) ahead of replication forks and preventing R loop formation. HDAC8 inactivation resulted in slowed fork progression and checkpoint kinase activation. Our findings indicate that HDAC8 guards the integrity of the replicating genome, and the cancer-specific synthetic lethality between HDAC8 and checkpoint kinases provides a promising replication stress-targeting strategy for treating a broad range of cancers.","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488297","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Antwi-Boasiako Oteng,Liu Liu,Yinghong Cui,Oksana Gavrilova,Huiyan Lu,Min Chen,Lee S Weinstein,Jonathan E Campbell,Jo E Lewis,Fiona M Gribble,Frank Reimann,Jürgen Wess
Following a meal, glucagon-like peptide-1 (GLP1) and glucose-dependent insulinotropic polypeptide (GIP), the two major incretins promoting insulin release, are secreted from specialized enteroendocrine cells (L- and K-cells, respectively). Although GIP is the dominant incretin in humans, the detailed molecular mechanisms governing its release remain to be explored. GIP secretion is regulated by the activity of G protein-coupled receptors (GPCRs) expressed by K-cells. GPCRs couple to one or more specific classes of heterotrimeric G proteins. In the present study, we focused on the potential metabolic roles of K-cell Gs. First, we generated a mouse model that allowed us to selectively stimulate K-cell Gs signaling. Second, we generated a mouse strain harboring an inactivating mutation of Gnas, the gene encoding the alpha-subunit of Gs, selectively in K-cells. Metabolic phenotyping studies showed that acute or chronic stimulation of K-cell Gs signaling greatly improved impaired glucose homeostasis in obese mice and in a mouse model of type 2 diabetes, due to enhanced GIP secretion. In contrast, K-cell-specific Gnas knockout mice displayed markedly reduced plasma GIP levels. These data strongly suggest that strategies aimed at enhancing K-cell Gs signaling may prove useful for the treatment of diabetes and related metabolic diseases.
进餐后，特化的肠内分泌细胞（分别为 L 细胞和 K 细胞）会分泌胰高血糖素样肽-1（GLP1）和葡萄糖依赖性促胰岛素多肽（GIP）这两种促进胰岛素释放的主要增量素。虽然 GIP 是人体中最主要的增量蛋白，但有关其释放的详细分子机制仍有待探索。GIP 的分泌受 K 细胞表达的 G 蛋白偶联受体（GPCR）活性的调节。GPCR 与一类或多类特定的异三聚体 G 蛋白偶联。在本研究中，我们重点研究了 K 细胞 Gs 的潜在代谢作用。首先，我们建立了一个小鼠模型，可以选择性地刺激 K 细胞 Gs 信号传导。其次，我们产生了一种小鼠品系，该品系在 K 细胞中选择性地携带 Gs α-亚基编码基因 Gnas 的失活突变。代谢表型研究表明，由于 GIP 分泌增强，急性或慢性刺激 K 细胞 Gs 信号大大改善了肥胖小鼠和 2 型糖尿病小鼠模型中受损的葡萄糖稳态。相反，K 细胞特异性 Gnas 基因敲除小鼠的血浆 GIP 水平明显下降。这些数据有力地表明，旨在增强 K 细胞 Gs 信号转导的策略可能被证明有助于治疗糖尿病和相关代谢疾病。
{"title":"Activation of Gs signaling in mouse enteroendocrine K-cells greatly improves obesity- and diabetes-related metabolic deficits.","authors":"Antwi-Boasiako Oteng,Liu Liu,Yinghong Cui,Oksana Gavrilova,Huiyan Lu,Min Chen,Lee S Weinstein,Jonathan E Campbell,Jo E Lewis,Fiona M Gribble,Frank Reimann,Jürgen Wess","doi":"10.1172/jci182325","DOIUrl":"https://doi.org/10.1172/jci182325","url":null,"abstract":"Following a meal, glucagon-like peptide-1 (GLP1) and glucose-dependent insulinotropic polypeptide (GIP), the two major incretins promoting insulin release, are secreted from specialized enteroendocrine cells (L- and K-cells, respectively). Although GIP is the dominant incretin in humans, the detailed molecular mechanisms governing its release remain to be explored. GIP secretion is regulated by the activity of G protein-coupled receptors (GPCRs) expressed by K-cells. GPCRs couple to one or more specific classes of heterotrimeric G proteins. In the present study, we focused on the potential metabolic roles of K-cell Gs. First, we generated a mouse model that allowed us to selectively stimulate K-cell Gs signaling. Second, we generated a mouse strain harboring an inactivating mutation of Gnas, the gene encoding the alpha-subunit of Gs, selectively in K-cells. Metabolic phenotyping studies showed that acute or chronic stimulation of K-cell Gs signaling greatly improved impaired glucose homeostasis in obese mice and in a mouse model of type 2 diabetes, due to enhanced GIP secretion. In contrast, K-cell-specific Gnas knockout mice displayed markedly reduced plasma GIP levels. These data strongly suggest that strategies aimed at enhancing K-cell Gs signaling may prove useful for the treatment of diabetes and related metabolic diseases.","PeriodicalId":520097,"journal":{"name":"The Journal of Clinical Investigation","volume":"60 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142488303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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