Pumilio RNA binding family member 1 deficiency activates anti-tumor immunity in hepatocellular carcinoma via restraining M2 macrophage polarization

doi:10.21203/rs.3.rs-3082586/v1

Download PDF

Research Article

Pumilio RNA binding family member 1 deficiency activates anti-tumor immunity in hepatocellular carcinoma via restraining M2 macrophage polarization

https://doi.org/10.21203/rs.3.rs-3082586/v1

This work is licensed under a CC BY 4.0 License

Journal Publication

published 24 May, 2024

Read the published version in Cell Cycle →

Version 1

posted

You are reading this latest preprint version

Pumilio RNA binding family member 1 (PUM1), which has been implicated in both the progression of colorectal cancer and the regulation of Inflammation, has not yet been associated with hepatocellular carcinoma PUM1 is essential for the transition of tumor-associated macrophages (TAMs) into the M2 polarization state. It does this by inhibiting anti-tumor immunity in hepatocellular carcinoma through a process mediated by TAMs that target CD8 + T cells, as demonstrated in this study using PUM1-knockout mice. By activating the cAMP signaling pathway, we have shown that PUM1 promotes the transformation of TAMs into pro-tumorigenic M2-like phenotypes. In order to emphasize the potential of PUM1 as an objective for immunotherapy centered on TAMs in the treatment of gastric carcinoma, the present investigation revealed the molecular mechanism underlying the pro-tumor role of PUM1 in this cancer.

Pumilio RNA binding family member 1 (PUM 1)

Tumor-associated macrophages (TAMs)

Tumor microenvironment (TME)

hepatocellular carcinoma (HCC)

Immunotherapy

The incidence and mortality of Hepatocellular carcinoma (HCC) rank fifth and third, respectively [1]. The majority of patients are identified at later stages, which generally results in an overall poor prognosis of the carcinoma. Moreover, Inflammation, multiple molecular occurrences, and several cellular signaling pathways were all linked to HCC advancement, apart from Hepatitis C virus and Hepatitis B virus infection [2]. Drug resistance, tumor recurrence, and metastasis significantly restrict the effectiveness of HCC treatments despite the availability of numerous treatment options, including targeted therapy and immunotherapeutic medications [3].

The tumor microenvironment (TME) has recently become recognized as having an essential part in the growth of tumors and could be a potential target for cancer therapy [4]. In the TME, tumor-associated macrophages (TAMs) are widely distributed and are regarded as crucial immunosuppressive cellular components [5]. The activation of TAMs is a complicated process that manifests itself in a variety of phenotypes during the progression of tumors [6]. Typically, early-stage tumor-initiating macrophages have an M1 phenotype and engage in anti-tumor actions [7], while the pro-tumorigenic TAMs receive instructions from the tumor cells to develop an immunosuppressive state and encourage the growth of the malignancy by either secreting a variety of anti-inflammatory chemicals or by blocking the cytotoxic activity of CD8 + T cells, thus evading the immune surveillance [8]. TAMs have anti-tumorigenic and pro-tumorigenic plasticity, due to which they are regarded as a potential target for therapeutic intervention against malignancies [9]. Several preclinical investigations have shown that preventing macrophage recruitment or modifying their characteristic phenotype to restricted M2 type might prevent tumor growth and enhance therapeutic response [10–14]. The identification of novel compounds capable of interfering with macrophages may provide novel targets for TAM-centered immunotherapy against cancer, and by possible reversal of the M2 state of TAMs in the tumors may thus be viewed as a potential therapeutic method.

Eukaryotic RNA binding proteins (RBPs) are highly conserved in nature. The PUF family, including the FBF and Pumilio, binds to the sequence regions post-transcriptionally, which are responsible for repressing gene expression. By binding targets specifically on the mRNAs at the 3'UTR region, the Pumilio-binding elements (PBEs) control cell differentiation and fate determination [15–17]. Recent genetic research on the genes that encode the RNA-binding proteins known as Pumilio-1 (Pum1) and Pumilio-2 (Pum2), members of the mammalian PUF family, have shown the functions of post-transcriptionally controlling growth, reproduction, and development of the eukaryotic brain in addition to malignant tumors progression [18–20]. An earlier study revealed that PUM1 is overexpressed in colon cancer cells that have developed Cetuximab resistance and is associated with TAM [21–22]. However, the mechanism of action of PUM1 in HCC is still largely unknown.

In the current study, we sought to understand better the processes by which PUM1 promotes the development of HCC by demonstrating that PUM1 inhibits anti-tumor immunity in HCC and is necessary for the M2 polarization of TAMs using PUM1-knockout mice. Our findings offer further proof showing the function of PUM1 in the tumor microenvironment in HCC.

Mice and Tumor Model

On a C57BL/6J background, wild-type (WT) and PUM1 knockout (PUM1-/-) mice were developed and acquired from Service Bio-Co., Ltd (Wuhan, China). The Experimental Animal Ethics Committee of Service Bio Co., Ltd (Wuhan, China) authorized the animal research and oversaw their execution according to guidelines. Female WT and PUM1-/- mice (6–8 weeks old) received subcutaneous injections of MHCC97H cells into the right flanks for subcutaneous tumor models. Tumor tissues were removed after the experiment was completed to do further examination. For the current study, we used bone marrow-derived macrophages (BMDMs). The C57BL/6J mice lost macrophages after being intraperitoneally injected with 200 mL of clodronate liposomes (YEASEN, Shanghai, China) 48 hours before the inoculations. Then, a combination of MHCC97H cells and macrophages obtained from WT or PUM1/mice was administered subcutaneously into the macrophage-depleted C57BL/6J mice. After measuring the diameter of the tumor, mice were executed according to protocol, and lymphocytes that entered the tumor were examined, as reported in prior work [23].

Cell Lines Used and Preparation of Tumor-Conditioned Medium

The Hep3B, HepG2, HCCLM3, Huh7, and MHCC-97H were the HCC cell lines under study. Moreover, the U937 and THP-1 were the two human monocyte cell lines used in the current work. All cell lines were taken and maintained following proper guidelines from the American Type Culture Collection (Maryland, USA). By treating both cell lines, human U937 cells and THP-1, to PMA (phorbol-12-myristate-13-acetate; 200 nmol/L, Sigma Aldrich, Taufkirchen, Germany) for 48 hours, the cells were transformed into mature macrophages. A complete growth medium was used to culture cancer cells, following its subsequent replacement by a serum-free medium when cells reached about 80% confluency. A fresh batch of conditioned medium (CM) was obtained after 24 hours, centrifuged for 5 minutes at 500g, filtered through 0.22 µm filters, and then stored at 80ºC.

Extraction and Stimulation of Mouse Primary Cells

As previously described, cells obtained from the bone marrow of female WT and PUM1-/- mice were cultured for 7 days in Iscove's modified Dulbecco's medium (Gibco, Grand Island, NY, USA) with FBS (concentration 10%) and M-CSF (50 ng/mL; PeproTech, NJ, USA) to obtain BMDMs [23]. BMDMs were cultured for 48 hours with IL4 / IL13 (20 ng/mL, PeproTech) to induce M2 macrophage differentiation. To obtain TAMs, a half volume of tumor CM was mixed with the culture medium being used, and the mixture was then allowed to incubate for a period of 48 hours.

In vitro phagocytosis assays

For this assay, several phenotypes of macrophages were added into 96-well low-attachment plates, and these plates were then co-cultured with Carboxyfluorescein succinimidyl ester (CFSE)-labeled MHCC-97H cells. All cells were collected after two hours, stained with F4/80 antibody, a murine macrophage surface marker, for 30 minutes on ice, and then subjected to flow cytometric analysis. The proportion of F4/ 80 + cells with fluorescence from CFSE was used to calculate the phagocytosis efficiency.

CD8 + T cell suppression assay

To achieve successful isolation of CD8 + T cells of the mouse from C57BL/ 6J mice, a CD8 + T cell isolation kit specific for the mouse was used that was manufactured by Stem Cell Technologies (Vancouver, Canada). After being stimulated with aCD3/ aCD28 and IL-2 (PeproTech) and co-cultured with various phenotypes of macrophages for a time period of 72 hours, CD8 + T cells supplemented with the eBioscience™ Cell Stimulation Cocktail (Invitrogen, Carlsbad, CA, USA) were incubated for 4 hours. The cells were then collected and subjected to flow cytometry analysis to determine whether they proliferated and expressed functional markers.

Statistical analysis

Statistical analysis was done by analysis of variance (ANOVA) or Student's t-tests e among the experimental groups. Whereas a p < 0.05 was statistically significant in all experiments. Statistical Package for the Social Sciences (SPSS) v17.0 (IBM, USA) and GraphPad Software (La Jolla, CA, USA) were used for all statistical analyses.

PUM1 deficient mice reduced the MHCC-97H tumor growth and polarized TAMs to M2-like phenotype

Subcutaneous injection of MHCC-97H cells in WT and PUM1 knockout mice was simultaneously done to explore the involvement of PUM1 in the tumor microenvironment. PCR testing verified that the mice had the expected genotype, and the PUM1 was in considerably low concentration in the primary BMDMs of PUM1-/- mice (Fig.S1A). We observed that the development of MHCC-97H tumors in PUM1-/- mice were noticeably slower than that of the WT group (Fig. 1A and B). The immune cell infiltration in the MHCC-97H tumors was investigated afterward. Tumors from PUM1-/- mice demonstrated reduced TAMs infiltration upon comparison to the tumors of WT mice. Despite this, there was no discernible variation in the frequency of all hematopoietic cells such as CD3e, CD11b, dendritic cells (DC), CD4 + T, and CD8 + T cells (Fig. 1C and D). More investigation showed that PUM1 deficiency suppressed CD206 + TAM cells (Fig. 1E) but increased the ratio of M1 to M2 macrophages and the frequency of IFN- γ + CD8 + T cells upon comparison to MHCC-97H-WT animal (Fig. 1F and G), suggesting that PUM1-/- mice had an attenuated M2-like immunosuppressive phenotype of TAMs. Although the difference was non-significant, we showed that the PUM1-/- mice had a lower ratio of monocytes in the blood (Fig.S1B). Additionally, PUM1-/- mice displayed lower blood serum levels of IL-10 (Fig.S1C), suggesting that the controlled phenotype change of TAMs to an M2-like state may help to prevent cancer formation in PUM1-/- animals.

PUM1 deficient macrophages are less polarized toward M2-like phenotypes

The effects of PUM1 deficiency on macrophages were then verified in vitro. Firstly, BMDMs were isolated and invigorated with IL-4 and IL-13 from PUM1-/- mice (PUM1-/-BMDMs) or WT mice (WT-BMDMs). PUM1-/- BMDMs had lower levels of ARG-1 and IL10, two typical M2 markers, compared to WT-BMDMs. In line with the current finding, the fraction of macrophages (CD206 + M2) was lower in PUM1-/-BMDMs (Fig. 2A and B). These findings suggest that PUM1 is necessary for polarization events in macrophages. For a better representation of the influence of PUM1 in the TME, WT-BMDMs were cultivated with CM isolated from the MHCC-97H cell culture (MHCC-97H-CM), which caused an upregulation of PUM1 in TAMs (Fig. 2C). The use of CM that was obtained from multiple human tumor cell cultures produced results that were comparable (Fig. 2D). In addition, we examined the differences in the state (phenotype) of BMDMs derived from wild-type and PUM1-/- animal. In comparison to their WT mouse counterparts, PUM1-/- BMDMs cultivated with MHCC-97H-CM showed enhanced gene expression, which was related to M1 type of macrophages (Fig. 2E) and reduced the expression of those genes involved in M2 type macrophages (Fig. 2F). Additional evidence suggested that PUM1-/- macrophages had less M2 polarization which was shown by flow cytometric analysis, and resulted in enhanced expression of CD86 and a decreased expression of CD206 (Fig. 2G and H).

Deficiency of PUM1 increases phagocytosis and decreases tumor-promoting activity

Next, we evaluated how PUM1 could impact macrophages' ability to phagocytose tumor cells. WT- or PUM1-/- BMDMs were co-cultured with CFSE-labeled MHCC-97H cells, respectively. Upon comparison to the WT mice, M2 macrophage from PUM1-/- mice showed enhanced phagocytosis of MHCC-97H cells (Fig. 3A). Similar outcomes were also seen in BMDMs treated with MHCC-97H-CM. When exposed to MHCC-97HCM, PUM1-/- BMDMs demonstrated increased macrophage phagocytosis than the WT-BMDMs. (Fig. 3B). Next, we explored how PUM1 affected tumor cell proliferation and migration with the assistance of macrophages. The CM boosted the capacity of MHCC-97H cell proliferation from WT macrophages. However, it was inhibited by CM from PUM1-/- macrophages upon comparison to WT-BMDMs. (Fig. 3C and D).

PUM1 deficiency limits the TAM-mediated inhibition of CD8 + T cells

TAMs have a suppressive effect on T-cell proliferation and the anti-tumor activity that accompanies it. Thus, we examined the immune modulatory effects of PUM1 in phagocytes by co-culturing M2 or CM-treated macrophages with CD8 + T cells. Co-culturing CD8 + T cells with M2 macrophages derived from PUM1-/- mice resulted in an increase in the ratio of activated CD8 + T cells to the total number of CD8 + T cells, as was proven by our research. This hypothesis was backed even further by the increase in CD8 + T cells that were positive for granzyme B, IFN-γ, or TNF alpha in comparison to the WT controls (Fig. 4A-C). CD8 + T cells with tumor CM-treated macrophages generated from PUM1-/- mice yielded similar findings. Following the co-culture technique with CM-treated PUM1-/- phagocytes, an apparent elevation of classic cytotoxic cytokines in CD8 + T cells were seen (Fig. 4D-F). These findings highlight PUM1's critical involvement in generating immunosuppressive TAM.

Activation of cAMP signaling by PUM1 induces macrophage M2 polarization

After that, we assessed the amount of cAMP produced and found that it increased in WT-BMDMs treated with IL4 / IL13 or MHCC-97H-CM but reduced in PUM1-/- BMDMs (Fig. 5A and B). In PUM1-/- macrophages, forskolin also significantly increased the quantity of IL-10 and ARG-1 (Fig. 5C–F). This data indicates PUM1 stimulating cAMP signaling to cause macrophage M2 polarization.

PUM1-/- BMDM inhibits MHCC-97H tumor growth and enhances antitumor immunity

Further, we wanted to evaluate whether PUM1-/- BMDMs will reconfigure TME and demonstrate anti-tumor effects in vivo. To examine this, MHCC-97H cells and PUM1-/- or WT BMDMs were combined before being implanted into mice with pre-depletion macrophages (C57BL/6J). PUM1-/- macrophage-containing MHCC-97H xenografts showed slower tumor development than those combined with WT macrophages (Fig. 6A and B). After this, the impact of PUM1-/- macrophages on the turnover of TAM and the infiltration of different immune cell groups, specifically CD8 + T cells in TME, was examined. The frequency of CD45 + cells (which are the hematopoietic cells) did not change, according to an analysis of immune cells infiltrating tumors. In the PUM1-/- macrophage co-cultured MHCC-97H group, TAMs tended to decline, although this trend did not achieve statistical significance (Fig. 6C and D). Contrarily, the number of CD86 + M1 type of macrophages rose, whereas the infiltration of CD206 + M2 type of macrophages significantly decreased (Fig. 6E and F). These findings imply that PUM1 transformed TAM into "anti-tumor" M1-like phenotypes from M2-like phenotypes which are known to cause tumor progression. The phenotypic change of TAMs was accompanied by tumors containing PUM1-/- macrophages displaying noticeably increased MHC class II on DC (Fig. 6G) and some of IFN-g + CD4 + T cells (Fig. 6H), showing increased activation of DC and responses was Th1-dominant. Additionally, the PUM1-/- macrophage group had enlarged granzyme B + CD8 + T cells (Fig. 6I) and IFN-γ + CD8 + T cells (Fig. 6J), with a decreased number of PD-1 + CD8 + T cells as shown in Fig. 6K. The results showed that PUM1-/- BMDM modified the immunosuppressive TME to improve anti-tumor immunity in the TME.

A significant portion of the TME comprises TAMs, which exhibit substantial plasticity in both function and phenotype in response to a wide variety of stimuli. Established tumors with TAMs are typically biased in the direction of the pro-tumorigenic M2 characters, encouraging tumor growth with reducing immunological anti-tumor response. Their presence tends to result in a poor prognosis in most malignancies [24]. TAMs have thus been regarded as a crucial aim for immunotherapy against cancers. In this study, we should identify an essential regulatory function of PUM1 regarding TAMs in the development of HCC, where PUM1 is critical for inducing an immunosuppressive phenotype in macrophages and blocking an anti-tumor immune response in TME. As a result, we conclude that PUM1 is a promising target for TAMs that aim to treat HCC using anti-tumor immunotherapy.

Our findings suggest that PUM1 was necessary for both the usual M2 polarization of macrophages and the induction of an immunosuppressive phenotype in macrophages in the tumor microenvironment (TME). Furthermore, PUM1 inhibition or knockdown in macrophages increased the number of the pro-inflammatory subtype of macrophages. This work explored PUM1's role in remodeling TAMs and the subsequent suppression of immunity in HCC. When compared to MHCC-97H-WT mice, our research discovered that PUM1-/- animals had reduced levels of CD206 + TAM and TAM, with higher levels of IFN-+ CD8 + T. These findings suggest that PUM1 had an immunosuppressive influence on the microenvironment of the developing tumor in addition to inhibiting the development of cancer. Additionally, our in vitro data showed that PUM1 encouraged TAMs to polarize into a pro-tumoral state. PUM1-/-macrophage inhibited tumor growth and improved immunity against cancer in vivo, as demonstrated by a sharp decline in the M2-like TAMs during tumor infiltration and an increase in anti-tumor lymphoid cell populations such as CD4 þ effector T cells, mature DC, and CD8 þ T cells. The immunological microenvironment of a carcinoma influences how various immunotherapeutic approaches will work [25]. Reprogramming immune cells and boosting anti-tumor immunity while overcoming treatment resistance caused by the microenvironment may be possible with the use of immunosuppressive TAMs and other similar agents. [26–29]. PUM1 might be a promising target for improving immunotherapies.

Various extracellular stimuli stimulate the second messenger cAMP through GPCRs [30–36]. Furthermore, cAMP and IL-4 work together synergistically to increase the expression of STAT6, and they may play the role of co-factors in reprogramming the macrophages. We further verified in the current study that cAMP contributed to the differentiation of TAMs in the TME via PUM1. Additionally, in PUM1-deficient macrophages, inducing cAMP synthesis restored M2-like behaviors. Our findings suggest that deficiency of PUM1 in TAMs prevents polarization of macrophages toward pro-tumor phenotypes with M2-like characteristics by reducing cAMP signaling.

Our findings demonstrate the significant regulatory function of PUM1 in TAMs. It enhances the immunosuppressive phenotype of macrophages, blocks the response of anti-tumor immunity, and also speeds up the growth of tumor in HCC. Because of this, PUM1 is a prospective pharmacological target for anti-tumor immunotherapy in HCC with a focus on TAM, and it may help in breaking the resistance against immune checkpoint blockade (ICB) therapies.

Conflict of Interest

The study had no possible conflicts of interest, according to the authors.

Funding

None

Acknowledgment

We sincerely thank American Editor (https://www.scimj.com/) for their professional linguistic services provided.

Author contributions

Liu Ouyang and Chen-ming Ni designed the research. All of the authors carried out the experiments, performed data analysis, wrote the manuscript. All of the authors revised the manuscript. All of the authors have read and approved the ﬁnal manuscript.

Xiao Y, Yang K, Liu P, Ma D, Lei P, Liu Q. Deoxyribonuclease 1-like 3 Inhibits Hepatocellular Carcinoma Progression by Inducing Apoptosis and Reprogramming Glucose Metabolism. Int J Biol Sci. 2022 Jan 1;18(1):82-95.
You Y, Wen D, Zeng L, Lu J, Xiao X, Chen Y, Song H, Liu Z. ALKBH5/MAP3K8 axis regulates PD-L1+ macrophage infiltration and promotes hepatocellular carcinoma progression. Int J Biol Sci. 2022 Aug 1;18(13):5001-5018.
Jin H, Qin S, He J, Xiao J, Li Q, Mao Y, Zhao L. New insights into checkpoint inhibitor immunotherapy and its combined therapies in hepatocellular carcinoma: from mechanisms to clinical trials. Int J Biol Sci. 2022 Mar 28;18(7):2775-2794.
Zhao Y, Guo S, Deng J, Shen J, Du F, Wu X, Chen Y, Li M, Chen M, Li X, Li W, Gu L, Sun Y, Wen Q, Li J, Xiao Z. VEGF/VEGFR-Targeted Therapy and Immunotherapy in Non-small Cell Lung Cancer: Targeting the Tumor Microenvironment. Int J Biol Sci. 2022 May 29;18(9):3845-3858.
Mo S, Shen X, Wang Y, Liu Y, Sugasawa T, Yang Z, Gu W, Nakajima T. Systematic single-cell dissecting reveals heterogeneous oncofetal reprogramming in the tumor microenvironment of gastric cancer. Hum Cell. 2023 Mar;36(2):689-701.
Zhang Z, Liang Z, Li D, Wang L, Chen Y, Liang Y, Jiao W, Niu H. Development of a CAFs-related gene signature to predict survival and drug response in bladder cancer. Hum Cell. 2022 Mar;35(2):649-664.
Katsuta E, Rashid OM, Takabe K. Clinical relevance of tumor microenvironment: immune cells, vessels, and mouse models. Hum Cell. 2020 Oct;33(4):930-937.
Tsukamoto H, Komohara Y, Oshiumi H. The role of macrophages in anti-tumor immune responses: pathological significance and potential as therapeutic targets. Hum Cell. 2021 Jul;34(4):1031-1039.
Kai K, Komohara Y, Esumi S, Fujiwara Y, Yamamoto T, Uekawa K, Ohta K, Takezaki T, Kuroda J, Shinojima N, Hamasaki T, Mukasa A. Macrophage/microglia-derived IL-1β induces glioblastoma growth via the STAT3/NF-κB pathway. Hum Cell. 2022 Jan;35(1):226-237.
Morito S, Kawasaki M, Nishiyama M, Sakumoto T, Hashiguchi M, Narita T, Kawaguchi A, Toda S, Aoki S. Microenvironmental elements singularity synergistically regulate the behavior and chemosensitivity of endometrioid carcinoma. Hum Cell. 2023 May;36(3):1147-1159.
Kersten K, Hu KH, Combes AJ, Samad B, Harwin T, Ray A, Rao AA, Cai E, Marchuk K, Artichoker J, Courau T, Shi Q, Belk J, Satpathy AT, Krummel MF. Spatiotemporal co-dependency between macrophages and exhausted CD8+ T cells in cancer. Cancer Cell. 2022 Jun 13;40(6):624-638.e9.
Kloosterman DJ, Akkari L. Macrophages at the interface of the co-evolving cancer ecosystem. Cell. 2023 Apr 13;186(8):1627-1651.
Wu G, Ma Z, Cheng Y, Hu W, Deng C, Jiang S, Li T, Chen F, Yang Y. Targeting Gas6/TAM in cancer cells and tumor microenvironment. Mol Cancer. 2018 Jan 31;17(1):20.
Wu Y, Zhang S, Gong X, Tam S, Xiao D, Liu S, Tao Y. The epigenetic regulators and metabolic changes in ferroptosis-associated cancer progression. Mol Cancer. 2020 Feb 27;19(1):39.
Yoon DS, Lee KM, Choi Y, Ko EA, Lee NH, Cho S, Park KH, Lee JH, Kim HW, Lee JW. TLR4 downregulation by the RNA-binding protein PUM1 alleviates cellular aging and osteoarthritis. Cell Death Differ. 2022 Jul;29(7):1364-1378.
Elagooz R, Dhara AR, Gott RM, Adams SE, White RA, Ghosh A, Ganguly S, Man Y, Owusu-Ansah A, Mian OY, Gurkan UA, Komar AA, Ramamoorthy M, Gnanapragasam MN. PUM1 mediates the posttranscriptional regulation of human fetal hemoglobin. Blood Adv. 2022 Dec 13;6(23):6016-6022.
Li X, Yang J, Chen X, Cao D, Xu EY. PUM1 represses CDKN1B translation and contributes to prostate cancer progression. J Biomed Res. 2021 Jul 16;35(5):371-382.
Wang Y, Sun L, Wang L, Yu H, Yu X, Zou Y. PUM1 modulates trophoblast cell proliferation and migration through LRP6. Biochem Cell Biol. 2021 Dec;99(6):735-740.
Gor R, Sampath SS, Lazer LM, Ramalingam S. RNA binding protein PUM1 promotes colon cancer cell proliferation and migration. Int J Biol Macromol. 2021 Mar 31; 174:549-561.
Murillo Carrasco A, Acosta O, Ponce J, Cotrina J, Aguilar A, Araujo J, Rebaza P, Pinto JA, Fujita R, Buleje J. PUM1 and RNase P genes as potential cell-free DNA markers in breast cancer. J Clin Lab Anal. 2021 Apr;35(4): e23720.
Yang Y, Su X, Shen K, Zhang C, Dai H, Ma H, Jiang Y, Shuai L, Liu Z, You J, Min K, Chen Z. PUM1 is upregulated by DNA methylation to suppress anti-tumor immunity and results in poor prognosis in pancreatic cancer. Transl Cancer Res. 2021 May;10(5):2153-2168.
Liu Q, Xin C, Chen Y, Yang J, Chen Y, Zhang W, Ye L. PUM1 Is Overexpressed in Colon Cancer Cells With Acquired Resistance to Cetuximab. Front Cell Dev Biol. 2021 Aug 10; 9:696558.
Dong F, Qin X, Wang B, Li Q, Hu J, Cheng X, Guo D, Cheng F, Fang C, Tan Y, Yan H, He Y, Sun X, Yuan Y, Liu H, Li T, Zhao Y, Kang C, Wu X. ALKBH5 Facilitates Hypoxia-Induced Paraspeckle Assembly and IL8 Secretion to Generate an Immunosuppressive Tumor Microenvironment. Cancer Res. 2021 Dec 1;81(23):5876-5888.
Li H, Luo F, Jiang X, Zhang W, Xiang T, Pan Q, Cai L, Zhao J, Weng D, Li Y, Dai Y, Sun F, Yang C, Huang Y, Yang J, Tang Y, Han Y, He M, Zhang Y, Song L, Xia JC. CircITGB6 promotes ovarian cancer cisplatin resistance by resetting tumor-associated macrophage polarization toward the M2 phenotype. J Immunother Cancer. 2022 Mar;10(3):e004029.
Lv B, Wang Y, Ma D, Cheng W, Liu J, Yong T, Chen H, Wang C. Immunotherapy: Reshape the Tumor Immune Microenvironment. Front Immunol. 2022 Jul 6;13:844142.
Xia L, Oyang L, Lin J, Tan S, Han Y, Wu N, Yi P, Tang L, Pan Q, Rao S, Liang J, Tang Y, Su M, Luo X, Yang Y, Shi Y, Wang H, Zhou Y, Liao Q. The cancer metabolic reprogramming and immune response. Mol Cancer. 2021 Feb 5;20(1):28.
Hinshaw DC, Shevde LA. The Tumor Microenvironment Innately Modulates Cancer Progression. Cancer Res. 2019 Sep 15;79(18):4557-4566.
Zhang Y, Mao Q, Xia Q, Cheng J, Huang Z, Li Y, Chen P, Yang J, Fan X, Liang Y, Lin H. Noncoding RNAs link metabolic reprogramming to immune microenvironment in cancers. J Hematol Oncol. 2021 Oct 15;14(1):169.
Ma G, Zhang Z, Li P, Zhang Z, Zeng M, Liang Z, Li D, Wang L, Chen Y, Liang Y, Niu H. Reprogramming of glutamine metabolism and its impact on immune response in the tumor microenvironment. Cell Commun Signal. 2022 Jul 27;20(1):114.
Gao Z, Lei WI, Lee LTO. The Role of Neuropeptide-Stimulated cAMP-EPACs Signalling in Cancer Cells. Molecules. 2022 Jan 5;27(1):311.
Lian J, Yue Y, Yu W, Zhang Y. Immunosenescence: a key player in cancer development.J Hematol Oncol. 2020 Nov 10;13(1):151.
Kumar N, Prasad P, Jash E, Saini M, Husain A, Goldman A, Sehrawat S. Insights into exchange factor directly activated by cAMP (EPAC) as potential target for cancer treatment. Mol Cell Biochem. 2018 Oct;447(1-2):77-92.
Gao Z, Lei WI, Lee LTO. The Role of Neuropeptide-Stimulated cAMP-EPACs Signalling in Cancer Cells. Molecules. 2022 Jan 5;27(1):311.
Desman G, Waintraub C, Zippin JH. Investigation of cAMP microdomains as a path to novel cancer diagnostics. Biochim Biophys Acta. 2014 Dec;1842(12 Pt B):2636-45.
Martiniuk A, Silva M, Amylon M, Barr R. Camp programs for children with cancer and their families: review of research progress over the past decade. Pediatr Blood Cancer. 2014 May;61(5):778-87.
Merkle D, Hoffmann R. Roles of cAMP and cAMP-dependent protein kinase in the progression of prostate cancer: cross-talk with the androgen receptor. Cell Signal. 2011 Mar;23(3):507-15.

supplementdata.docx

Download PDF

Journal Publication

published 24 May, 2024

Read the published version in Cell Cycle →

Version 1

posted

You are reading this latest preprint version

Pumilio RNA binding family member 1 deficiency activates anti-tumor immunity in hepatocellular carcinoma via restraining M2 macrophage polarization

Status:

Journal Publication

Version 1

Abstract

Figures

Background

Material and methods

Mice and Tumor Model

Cell Lines Used and Preparation of Tumor-Conditioned Medium

Extraction and Stimulation of Mouse Primary Cells

CD8 + T cell suppression assay

Statistical analysis

Results

PUM1 deficient mice reduced the MHCC-97H tumor growth and polarized TAMs to M2-like phenotype

PUM1 deficient macrophages are less polarized toward M2-like phenotypes

Deficiency of PUM1 increases phagocytosis and decreases tumor-promoting activity

PUM1 deficiency limits the TAM-mediated inhibition of CD8 + T cells

Activation of cAMP signaling by PUM1 induces macrophage M2 polarization

PUM1-/- BMDM inhibits MHCC-97H tumor growth and enhances antitumor immunity

Discussion

Conclusions

Declarations

References

Supplementary Files

Status:

Journal Publication

Version 1