CAR-T cells costimulated with 4-1BB or ICOS persist in xenograft models but those constructed with CD28 exhibit rapid clearance. tains a single disulphide-linked IgV-like domain. A first signal, which is antigen-specific, is provided through the T cell receptor (TCR) which interacts with peptide-MHC molecules on the membrane of antigen presenting cells (APC). In another direction with accessory proteins, Zhao et al. Would you like email updates of new search results? Willemsen RA, Ronteltap C, Chames P, Debets R, Bolhuis RL. We and others have shown that CARs with a CD28 co-stimulatory domain drive high levels of T cell activation that also lead to exhaustion and shortened persistence. T cells require both primary and costimulatory signals for optimal activation. CARs containing the ICOS costimulatory domain have yet to be evaluated clinically; however, mesothelin-specific ICOS-based CAR-T cells demonstrate robust persistence and antitumor efficacy in … 1992; Azuma et al. Ralph Willemsen. The costimulatory receptors CD28, CTLA4, ICOS and PD1 are composed of single extracellular IgV-like domains, whereas BTLA has one IgC-like domain. An obstacle to the development of chimeric antigen receptor (CAR) T cells is the limited understanding of CAR T-cell biology and the mechanisms behind their antitumor activity. This suggests that mut06 CAR T cells are more resistant to exhaustion. To examine exhaustion, we ex vivo stimulated CAR T cells with target cells expressing CD19 and PDL1 and found mut06 CAR T cells had increased IFNγ (42%), TNFα (62%) and IL2 (73%) secretion compared to exhausted non-mutated CD28 CAR T cells. Clin Transl Immunology. Coronavirus: Find the latest articles and preprints Sign in or create an account. Co-stimulation is a secondary signal which immune cells rely on to activate an immune response in the presence of an antigen-presenting cell. We found that most of the exhaustion related genes were upregulated in m1928z CAR T cells while they were decreased in m19hBBz. Curr Mol Med. CD28 and cytotoxic T lymphocyte antigen-4 (CTLA4) homologs have been recently identified in rainbow trout. The proliferative response induced by anti-CD28 alone in CD3 − CAR + T cells was similar in CAR T cells with a 4-1BB or a CD28 costimulatory domain in their ICD (Supplementary Figure 4A). Enhanced cytokine release (e.g. The CD28 Costimulatory Domain Enhances Antigen-Specific Cytotoxicity and Cytokine Production1 Ralph A. Willemsen2,* Cees Ronteltap,* Patrick Chames,† Reno Debets,* and Reinder L. H. Bolhuis* T cells require both primary and costimulatory signals for optimal activation. 1989; Balzano et al. The primary Ag-specific signal is delivered by engagement of the TCR. The binding domain for JCAR015 is SJ25C1 and the costimulatory domain is CD28, whereas JCAR017 has a binding domain of FMC63 and a costimulatory domain of 4-1BB. Ralph Willemsen. Li:ImmuneBro Therapeutics: Other: sole shareholder . 1993; Freeman et al. All of these costimulatory factors play important roles in the activation and function of T cells. The extracellular domain of CD28 binds to B7 proteins using a MYPPPY motif and this interaction initiates the costimulatory signal transduction cascade (Freeman et al. Using the same system, the authors also showed that there was a decrease in the recruitment of the PI3K SH2 domain to CD28. The CD28 domain includes 3 intracellular subdomains (YMNM, PRRP, and PYAP) that regulate signaling pathways post TCR-stimulation, but it is unknown how they modulate activation and/or exhaustion of … We and others have shown that CARs with a CD28 co-stimulatory domain drive high levels of T cell activation that also lead to exhaustion and shortened persistence. The mut06 CAR T cell gene expression pattern was more similar to m19hBBz with exhaustion related genes downregulated compared to m1928z (Fig 1D). Dobbins et al . Here, we show that a single amino acid residue in CD28 drove T cell exhaustion and hindered the … ICOS, a T cell costimulatory receptor of the B7-CD28 superfamily, which also includes inhibitory receptors CTLA-4 and PD-1, shares the signaling motif YMXM with CD28 in its intracellular domain. 1993; Freeman et al. 1989; Balzano et al. Selecting costimulatory domains for chimeric antigen receptors: functional and clinical considerations. Disclosed herein are chimeric antigen receptor (CAR) polypeptides, which can be used with adoptive cell transfer to target and kill cancers, that comprise a co-stimulatory signaling region having a mutated form of a cytoplasmic domain of CD28 that enhances CAR-T cell function, e.g. With CD33 CD28 against THP1, there was clearance in compartments bone marrow, however CD33 CD28 CAR was not able to prevent the development of EMD. T cells require two signals to become fully activated. Savoldo B, Ramos CA, Liu E, Mims MP, Keating MJ, Carrum G, Kamble RT, Bollard CM, Gee AP, Mei Z, Liu H, Grilley B, Rooney CM, Heslop HE, Brenner MK, Dotti G. J Clin Invest. In vivo, we found that mice given CAR T cells with only a PYAP CD28 endodomain had a significant survival advantage, with 100% of mice alive after 62 days compared with 50% for mice with an unmutated endodomain. Involved in T-cell activation, the induction of cell proliferation and cytokine production and promotion of T-cell survival. After 24hr stimulation with CD19 target cells mut06 CAR T cells had a significant reduction in pAkt compared to m1928z CAR T cells, which is a critical signaling mediator in the NFAT and NR4A1 transcription factor pathways. Molecules involved in signal transduction in activated T cells (modified from ALEGRE1) 170 D. Boc´ko et al. To examine differences in the accessibility of exhaustion related genes we performed ATAC-seq and found NFAT (Nfatc1) and NR4A2 (Nr4a2) had lower chromatin accessibility profiles in mut06 compared to m1928z (Fig 1E). CD28 and cytotoxic T lymphocyte antigen-4 (CTLA4) homologs have been recently identified in rainbow trout. (A) Designs of five CAR against CD19 bearing a 4-1BB costimulatory domain and differing by their hinge domain (HD) and their transmembrane domain (TMD). Superantigens—bacterial virulence factors—cause toxic shock by hyperinducing inflammatory cytokines. CD28 is the most effective and well-characterized T-cell costimulatory receptor discovered to date. Here we show that CD28-B7-mediated interactions are likely involved in the relocalization of Tec at the contact zone between T cells and APC. CD28 co-stimulatory domain-targeted mutations enhance chimeric antigen receptor T-cell function CD28 co-stimulatory domain-targeted mutations enhance chimeric antigen receptor T-cell function Cancer Immunol Res. eCollection 2020 Sep 25. CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatory signals required for T cell activation and survival. JCAR017 contains an ablative technology to provide better control of proliferation and survival of the engineered T cells, which is not included in JCAR015. CD28 family of costimulatory receptors is comprised of molecules with a single V-type extracellular Ig domain, a transmembrane and an intracytoplasmic region with signaling motifs. T umor-associated Ags (1) or MHC class I-presented tumor rejection Ags (TRA) 3 (2,3) can be targeted by CTL-expressing (chimeric) single chain (sc) or two chain mAbs (4 -10) … The cytoplasmic domain of DAP10 has one known signaling motif, a YINM-sequence, that is also found in the CD28 costimulatory receptor [8,9]. doi: 10.1158/2326-6066.CIR … Patrick Chames. CD28 includes at least three intracellular sub-domains (YMNM, PRRP, and PYAP) that regulate costimulation post–T-cell receptor (TCR) stimulation, but it is unknown how each This site needs JavaScript to work properly. The first-generation CAR contained only the CD3ζ domain, which was designated Z. Second-generation CARs also contained costimulatory domains. teristics: a CD28 costimulatory domain stimulates more powerful cytotoxic ability of CAR-T cells, whereas the 4-1BB and ICOS costimulatory domain induces longer persistence of CAR-Tcells [9]. This included the 2nd generation m1928z CAR, which contains the CD28 costimulatory domain (FIG. In some embodiments, the mutated form reduces CAR-T cell exhaustion. This work allows for the development of enhanced CAR T-cell therapies by optimizing CAR T-cell costimulation. demonstrated that a second-generation CAR containing a CD28 costimulatory domain with CD3 ζ, expressed together with 4-1BBLigand in trans (CD28z/41BBL), induced a more potent antitumor effect than a third-generation CAR containing both CD28 and 4-1BB in cis . This would result in the kinase-dependent phosphorylation of the potential PI3-kinase, GRB-2, and ITK binding site (C). https://orcid.org. The genes encoding CD28 costimulatory receptor are found on chromosome 1 in mice and chromosome 2 in humans. In this study, we confirmed that the CD28 costimulatory domain-containing PD-L1 CCR co-stimulated and promoted the activation of CAR-T cells, especially for CD4 + CAR-T cells. In addition to signaling by the T cell receptor (TCR), signaling by the costimulatory receptor CD28 is required for full activation of naïve T cells and the generation of regulatory T (Treg) cells. Differential signaling through the CAR costimulatory domain can alter the T cell metabolism, memory differentiation, and influence long-term persistence. A detailed understanding of the mechanism of CD28-dependent exhaustion in CAR T cells will allow the design of a CAR less prone to exhaustion and reduce relapse rates. Mouse and human CD28 molecules share approximately 68% amino acid identity. CD8α, CD3ζ T cell activation domain and a costimulatory molecule CD28. Bethesda, MD 20894, Copyright Other members of the subfamily include ICOS, CTLA4, PD1, PD1H, TIGIT, and BTLA ( Chen and Flies, 2013 , Anderson et al., 2016 in this issue). 2020 Nov 13;canimm.0253.2020. 2019 Dec 1;25(23):7126-7138. doi: 10.1158/1078-0432.CCR-19-0421. In vitro, we found mutated CAR T cells with only a functional PYAP (mut06) subdomain secrete significantly less IFNγ, IL6, and TNFα after 24hr stimulation compared to non-mutated CD28 CAR T cells, but greater than the 1st generation m19z CAR. We also found that exhaustion related genes Havcr2 (TIM3), Pdcd1 (PD1), and Lag3 (LAG3) all had greatly reduced chromatin accessibility in mut06 CAR T cells compared m1928z. The CD28 domain includes 3 intracellular subdomains (YMNM, PRRP, and PYAP) that regulate signaling pathways post TCR-stimulation, but it is unknown how they modulate activation and/or exhaustion of CAR T cells. A detailed understanding of the mechanism of CD28-dependent exhaustion in CAR T cells will allow the design of a CAR less prone to exhaustion and reduce relapse rates. Combined CD28 and 4-1BB Costimulation Potentiates Affinity-tuned Chimeric Antigen Receptor-engineered T Cells. The most frequently used costimulatory domains derive from the CD28 family (CD28 and ICOS, Inducible T Cell Costimulator) and the tumor necrosis factor receptor (TNFR) family (4-1BB, CD27, and OX40). : CD28 Costimulatory Molecule FIGURE 4. During the activation of lymphocytes, co-stimulation is often crucial to the development of an effective immune response. Kymriah and Yescarta use the same scFv, which recognizes the B cell antigen CD19, but Yescarta incorporates a CD28-derived costimulatory domain, whereas Kymriah incorporates a 4-1BB domain … 1. By generating CAR constructs consisting of genes encoding single-chain antibodies (scFv), costimulatory domains (CD28 or TNFRSF9), and CD247 signaling domains for T cell proliferation and activation, T cells have the ability to attack tumor cells . CD28 costimulatory domain protects against tonic signaling-induced functional impairment in CAR-Tregs. The extracellular domain of CD28 binds to B7 proteins using a MYPPPY motif and this interaction initiates the costimulatory signal transduction cascade (Freeman et al. Justin C. Boucher, Gongbo Li, Hiroshi Kotani, Maria Cabral, Dylan Morrissey, Sae Bom Lee, Kristen Spitler, Nolan Beatty, Bishwas Shrestha, Bin Yu, Aslamuzzaman Kazi, Xuefeng Wang, Said Sebti, Marco L. Davila; Mutation of the CD28 Co-Stimulatory Domain Confers Enhanced CAR T Cell Function. Xu X, Huang W, Heczey A, Liu D, Guo L, Wood M, Jin J, Courtney AN, Liu B, Di Pierro EJ, Hicks J, Barragan GA, Ngai H, Chen Y, Savoldo B, Dotti G, Metelitsa LS. Clipboard, Search History, and several other advanced features are temporarily unavailable. To determine how optimized CAR signaling affected T cell exhaustion we looked at 22 genes that are upregulated when NFAT is constitutively active and overlap with genes identified as important for T cell exhaustion. 8600 Rockville Pike Epub 2019 Sep 4. T cell retargeting with MHC class I-restricted antibodies: the CD28 costimulatory domain enhances antigen-specific cytotoxicity and cytokine production Publication Publication. J Immunol 2005; 174: 7853 –8. CAR-T cells are activated by recognizing TAA by scFv on the surface of T cells, and then activate the intracellular signaling domains connected by scFv to induce downstream signaling pathways involving T cell proliferation, activation and cytokine production. 2019 May 11;8(5):e1049. FOIA Other sequences similar to mammalian CD28 family CD28 is the most effective and well-characterized T-cell costimulatory receptor discovered to date. The CD28 domain includes 3 intracellular subdomains (YMNM, PRRP, and PYAP) that regulate signaling pathways post TCR-stimulation, but it is unknown how they modulate activation and/or exhaustion of CAR T cells. Patrick Chames. Route of 41BB/41BBL Costimulation Determines Effector Function of B7-H3-CAR.CD28ζ T Cells. CD28 family of costimulatory receptors is comprised of molecules with a single V-type extracellular Ig domain, a transmembrane and an intracytoplasmic region with signaling motifs. Please enable it to take advantage of the complete set of features! ©2020 American Association for Cancer Research. As principal costimulatory receptor, CD28 is a critical regulator of the immune response (1–3). CD28 Costimulatory Molecule – Expression, Structure and Function D. Boc´ko et al. We observed that mutant CAR T cells remained more sensitive to antigen after ex vivo antigen and PD-L1 stimulation, as demonstrated by increased cytokine production. These experiments suggest that although the CD28 costimulatory domain is more potent than 4-1BB in Molm14, the potency of CD28 is still not able to overcome EMD in all models. The conventional or costimulatory binding of B7-2 to CD28 results in the liberation of the carboxylterminus of CD28, making this proline-rich domain available for SH3 domain interaction. We and others have shown that CARs with a CD28 costimulatory domain drive high T-cell activation, which leads to exhaustion and shortened persistence. Latest research progress of allogeneic CAR-T cell therapy. The CD28 co-stimulatory domain used in CAR T cells is from the cytoplasmic tail of CD28, which has no intrinsic enzymatic activity but contains subdomains or motifs that regulate T cell signaling. COVID-19 is an emerging, rapidly evolving situation. Tec, the prototypical member of this kinase family, can interact with CD28, which is a costimulatory molecule. The mutant CAR T cells also had a reduction of exhaustion-related transcription factors and genes such as Nfatc1, Nr42a, and Pdcd1 Our results demonstrated that CAR T cells with a mutant CD28 endodomain have better survival and function. Unable to load your collection due to an error, Unable to load your delegates due to an error. Accessibility Different costimulatory domains endow CAR-T cells with different characteristics: a CD28 costimulatory domain stimulates more powerful cytotoxic ability of CAR-T cells, whereas the 4-1BB and ICOS costimulatory domain induces longer persistence of CAR-T cells .

A combinatorial costimulatory design allows the use of very low-affinity binding domains (K _d < 1 μmol/L) for the construction of safe but … 1. Menu. T Cell Retargeting with MHC Class I-Restricted Antibodies: The CD28 Costimulatory Domain Enhances Antigen-Specific Cytotoxicity and Cytokine Production. 1993; Rudd and Schneider 2003). domain can be significantly improved when a CD28 costimulatory domain is used. Weinkove R, George P, Dasyam N, McLellan AD. In the case of T cells, two stimuli are required to fully activate their immune response. An obstacle with continued clinical development of CAR T cells is the limited understanding of their biology and mechanisms of anti-tumor immunity. Davila:Atara: Research Funding; Celgene: Research Funding; GlaxoSmithKline: Consultancy; Novartis: Research Funding; Anixa: Consultancy; Bellicum: Consultancy; Adaptive: Consultancy; Precision Biosciences: Consultancy. 2013 Aug;13(7):1079-88. doi: 10.2174/1566524011313070003. 2020 Jun 23;18:202-214. doi: 10.1016/j.omto.2020.06.018. P30 CA076292/CA/NCI NIH HHS/United States, NCI CPTC Antibody Characterization Program. The primary Ag-specific signal is delivered by engagement of the TCR. by reducing CAR-T cell exhaustion. To find a mechanistic explanation for this observation we examined CAR T cell signaling. These studies suggest that the type of costimulatory domain or quality of signaling … We and others have shown that CARs with a CD28 co-stimulatory domain drive high levels of T cell activation causing acute toxicities, but also lead to T cell exhaustion and shortened persistence. doi: 10.1002/cti2.1049. Also, cytotoxicity was enhanced compared to non-mutated CARs (Fig 1B). The primary Ag-specific signal is delivered by engagement of the TCR. ScFv receptors comprising CD28 mediate a more potent cytolytic T cell response. Mouse and human CD28 molecules share approximately 68% amino acid identity. A second signal, the co-stimulatory signal, is antigen nonspecific and is provided by the interaction between co-stimulatory molecules expressed on the membrane of APC and the T cell. Ligation of CD28 recruits the phosphatidylinositol 3‐kinase (PI3K), the adapter Grb‐2, the serine/threonine... 4‐1BB (CD137). 1992; Azuma et al. Using a pre-clinical immunocompetent mouse tumor model, we found the mut06 CAR T cell treated mice had a significant survival advantage compared to non-mutated CD28 CAR T cells (Fig 1C). Epub 2011 Apr 11. All of these costimulatory factors play important roles in the activation and function of T cells. Europe PMC. exhaustion in CAR T cells with a CD28 endodomain is induced by specific signaling pathways and transcription factors and not just the strength of signaling. The choice of costimulatory domain influences the proliferation rate of IIICAR.GD2 T cells upon extended in vitro culture Our initial results showed no significant differences in terms of cytotoxic and anti-tumor activities between IICAR.GD2 (including as costimulatory molecule either CD28, or OX40 Isoform 3 enhances CD40L-mediated activation of NF-kappa-B and kinases MAPK8 and PAK2 in T-cells (PubMed:15067037). Results: We show that the inferior cytotoxicity and cytokine secretion mediated by CD38 CARs of very low–affinity (K d < 1.9 × 10 −6 mol/L) bearing a 4-1BB intracellular domain can be significantly improved when a CD28 costimulatory domain is used. CAR-T cells bearing a 4-1BB costimulatory endodomain seem to persist for longer time in patient circulation in comparison with CAR-T cells having a CD28 costimulatory domain (3, 5, 20), due to better maintenance of a memory phenotype and reliance on oxidative metabolism (21–24). National Library of Medicine CD28 is the founding member of a subfamily of costimulatory molecules characterized by an extracellular variable immunoglobulin-like domain. Additionally, mut06 had decreased p-NFAT compared to m1928z when examined by western blot. Different costimulatory domains endow CAR-T cells with different characteristics: a CD28 costimulatory domain stimulates more powerful cytotoxic ability of CAR-T cells, whereas the 4-1BB and ICOS costimulatory domain induces longer persistence of CAR-T cells . T cell stimulation through CD28 in addition to the T-cell receptor (TCR) can provide a potent signal for the production of various interleukins (IL-6 in particular). Privacy, Help Superantigens must bind CD28 to elicit cytokine overexpression through a hitherto unknown mechanism. costimulatory receptor B7-2 dimer interface As principal costimulatory receptor, CD28 is a critical regulator of the immune response (1 ⇓ – 3). About. This work led us to hypothesize that by incorporating null mutations of CD28 subdomains (YMNM, PRRP, or PYAP), we could optimize CAR T-cell costimulation and enhance function. Receptors CTLA4, CD28 and ICOS are covalent homodimers, due to an interchain disulphide linkage. T-cell activation is mediated both by antigen and by interaction between principal costimulatory receptors B7-2 and CD28. Expressed constitutively on T cells, CD28 is a homodimer that interacts with its B7 coligands, transducing the signal essential for an immediate T-cell response (2–5). Enhances the production of IL4 and IL10 in T-cells in conjunction with TCR/CD3 ligation and CD40L costimulation (PubMed:8617933). Expressed constitutively on T cells, CD28 is a homodimer that interacts with its B7 coligands, transducing the signal essential for an immediate T-cell response (2 ⇓⇓ – 5). ICOS, a T cell costimulatory receptor of the B7-CD28 superfamily, which also includes inhibitory receptors CTLA-4 and PD-1, shares the signaling … 2011 May;121(5):1822-6. doi: 10.1172/JCI46110. This led us to hypothesize that by incorporating null mutations of CD28 subdomains (Fig 1A) we could optimize CAR T cell signaling and reduce exhaustion. The costimulatory ligands B71, B72, B7H2, B7H1 and B7DC, have a membrane proximal IgC-like domain and a membrane distal IgV-like domain that is responsible for receptor binding and dimerization. Moreover, anti-CD28 induced the secretion of multiple cytokines by CD3 − CAR + T cells but not by CD3 + CAR − or CD3 − CAR − control cells (Supplementary Figure 4B). Search for other works by this author on: © 2019 by The American Society of Hematology, Copyright ©2020 by American Society of Hematology, 703.Adoptive Immunotherapy: Mechanisms and New Approaches, https://doi.org/10.1182/blood-2019-122954. T cell retargeting with MHC class I-restricted antibodies: the CD28 costimulatory domain enhances antigen-specific cytotoxicity and cytokine production. Moreover, anti-CD28 induced the secretion of multiple cytokines by CD3 − CAR + T cells but not by CD3 + CAR − or CD3 − CAR − control cells (Supplementary Figure 4B). Title: CD80-Fc Fusion Protein FPT155 Definition: A recombinant fusion protein composed of the extracellular domain (ECD) of human CD80 (B7.1) fused to a human immunoglobulin G1 IL-2) and proliferation were found in dual-targeted CAR-T cells expressing a suboptimal CD19/HER2 CAR and PD-L1 CCR, which is consistent with a previous study that showed the CD28 … Clin Cancer Res. 1). The extracellular tail of the CD28 receptor possesses the Fig. ICOS, a T cell costimulatory receptor of the B7-CD28 superfamily, which also includes inhibitory receptors CTLA-4 and PD-1, shares the signaling motif YMXM with CD28 in its intracellular domain. CD28 costimulation improves expansion and persistence of chimeric antigen receptor-modified T cells in lymphoma patients. Receptors CTLA4, CD28 and ICOS are covalent homodimers, due to an interchain disulphide linkage. The cytoplasmic domain of CD28 is both necessary and sufficient for costimulation of interleukin-2 secretion and association with phosphatidylinositol 3'-kinase. The disclosed CARs comprise a costimulatory signaling region comprising a mutated form of the cytoplasmic domain of CD28 that enhances CAR-T cell function. Journal of Immunology . CAR with the T-cell costimulatory domain CD28 was designated CD28.Z, and CAR with NK-cell-associated costimulatory domain DNAM1 or 2B4 was designated DNAM1.Z and 2B4.Z, respectively. : CD28 Costimulatory Molecule DOROTA BOC´KO1, AGATA KO SMACZEWSKA1, LIDIA CISZAK1, RENATA TEODOROWSKA1 and IRENA FRYDECKA1, 2* 1 Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Po land, 2 Department of Hematology, Medical University, Wrocław, Poland A bstract. Costimulatory domains within chimeric antigen receptors CD28. Blood 2019; 134 (Supplement_1): 248. doi: https://doi.org/10.1182/blood-2019-122954. The proliferative response induced by anti-CD28 alone in CD3 − CAR + T cells was similar in CAR T cells with a 4-1BB or a CD28 costimulatory domain in their ICD (Supplementary Figure 4A). Collectively, these results show that … Nguyen P, Okeke E, Clay M, Haydar D, Justice J, O'Reilly C, Pruett-Miller S, Papizan J, Moore J, Zhou S, Throm R, Krenciute G, Gottschalk S, DeRenzo C. Mol Ther Oncolytics. Differential signaling through the CAR costimulatory domain can alter the T cell metabolism, memory differentiation, and influence long-term persistence. Kymriah and Yescarta use the same scFv, which recognizes the B cell antigen CD19, but Yescarta incorporates a CD28-derived costimulatory domain, whereas Kymriah incorporates a 4-1BB domain (the two constructs also have different hinge and transmembrane regions). rbtCD28 and rbtCTLA4 are composed of an extracellular Ig-superfamily V domain, a transmembrane region, and a cytoplasmic tail.

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